Information on EC 6.1.1.4 - leucine-tRNA ligase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota, Archaea

EC NUMBER
COMMENTARY
6.1.1.4
-
RECOMMENDED NAME
GeneOntology No.
leucine-tRNA ligase
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
bi uni uni bi ping-pong mechanism with ordered addition of ATP and leucine and random release of AMP and leucyl-tRNALeu
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
each amino acid is bound through its carboxyl group to the terminal nucleotide (2'- or 3'-hydroxyl end) of specific polynucleotide chains
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
ATP and tRNA are bound to the enzyme in almost random order, and diphosphate is dissociated before the rate-limiting step
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
A293 is important for the stability of the enzyme conformation and the editing function and probably is involved in the ATP binding
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
active site structure and mechanism, the editing active site binds the two different substrates using a single amino acid discriminatory pocket while preserving he same mode of adenine recognition, Asp347 is involved in the editing process, mechanism of hydrolysis
Q72GM3
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Asp345 is involved in the editing process, mechanism of hydrolysis
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Asp419 is involved in the editing process, mechanism of hydrolysis
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
recognition of the tRNALeu requires the discriminator base A73 and the long variable arm of appropriate stem length, especially the Haloferax volcanii-specific loop sequence A47CG47D and U47H at the base of the helix
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
residue E292 is important for aminoacylation activity
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Aminoacylation
-
-
-
-
esterification
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Aminoacyl-tRNA biosynthesis
-
-
leucine metabolism
-
-
tRNA charging
-
-
SYSTEMATIC NAME
IUBMB Comments
L-leucine:tRNALeu ligase (AMP-forming)
-
SYNONYMS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Leucine translase
-
-
-
-
Leucine--tRNA ligase
-
-
-
-
Leucyl-transfer ribonucleate synthetase
-
-
-
-
Leucyl-transfer ribonucleic acid synthetase
-
-
-
-
Leucyl-transfer RNA synthetase
-
-
-
-
Leucyl-tRNA synthetase
-
-
-
-
LeuRS
-
-
-
-
Synthetase, leucyl-transfer ribonucleate
-
-
-
-
CAS REGISTRY NUMBER
COMMENTARY
9031-15-6
-
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
class I enzyme
-
-
Manually annotated by BRENDA team
class I enzyme, 2 genes leuS and leuS'
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
high-MW enzyme complex; temperature-sensitive mutant ts025C1; wild-type
-
-
Manually annotated by BRENDA team
high-MW enzyme complex; temperature-sensitive mutant tsH1; wild-type
-
-
Manually annotated by BRENDA team
Torulopsis
-
-
Manually annotated by BRENDA team
Cyberlindnera jadinii Torulopsis
Torulopsis
-
-
Manually annotated by BRENDA team
B or ML30
-
-
Manually annotated by BRENDA team
class I enzyme
-
-
Manually annotated by BRENDA team
gene leuS
-
-
Manually annotated by BRENDA team
K12 strain AE52, a leucine-auxotrophic strain
-
-
Manually annotated by BRENDA team
MRE 600
-
-
Manually annotated by BRENDA team
Escherichia coli MRE 600
MRE 600
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
class I enzyme
SwissProt
Manually annotated by BRENDA team
gene LARS1
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
wild-type and temperature-sensitive and leucine-auxotroph mutant leu-5
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
respiratory deficient mutants. The phenotype is a consequence of a mutation in a nuclear gene coding for mitochondrial leucyl-tRNA synthetase
-
-
Manually annotated by BRENDA team
purified recombinant enzyme
Uniprot
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
-
the family of leucyl-tRNA synthetases is divided into prokaryotic and eukaryal/archaeal groups according to the presence and position of specific insertions and extensions. e.g. the LSD1, i.e. leucine-specific domain 1, which is naturally present in eukaryal/archaeal LeuRSs, but absent from prokaryotic LeuRSs. The LSD1s from organisms of both groups are dispensable for post-transfer editing
malfunction
-
replacement of Giardia lamblia eukarya-specific insertion 1, GlESI, by human eukarya-specific insertion 1, HsESI, impairs leucine activation, aminoacylation and post-transfer editing functions without changing the editing specificity
metabolism
-
isoform LeuRS1 generates Leu-tRNALeu for protein biosynthesis and exhibits obvious post-transfer editing activity to prevent generation of mischarged tRNALeu
physiological function
-
existence of a tRNA-independent pretransfer editing pathway in leucyltRNA synthetases from Aquifex aeolicus. This editing pathway is distinct from the post-transfer editing site and may occur at the synthetic catalytic site
physiological function
-
leucyl-tRNA synthetase is an essential RNA splicing factor for yeast mitochondrial introns. RNA deletion mutants of the large bI4 intron are active in RNA splicing and the activity of the minimized bI4 intron is enhanced in vitro by the presence of the bI4 maturase or LeuRS
physiological function
-
the A3243G mutation of the tRNALeu gene causes mitochondrial encephalomyopathy, lactic acidosis, and stroke-like symptoms and 2% of cases of type 2 diabetes. The alteration of aminoacylation of tRNALeu(UUR) caused by the A3243G mutation leads to mitochondrial translational defects and thereby reduces the aminoacylating efficiencies of tRNALeu(UUR) as well as of tRNAAla and tRNAMet
physiological function
-
aminoacyl-tRNA synthetases are critical for the translational process, catalyzing the attachment of specific amino acids to their cognate tRNAs. To ensure formation of the correct aminoacyl-tRNA, and thereby enhance the reliability of translation, several aminoacyl-tRNA synthetases have an editing capability that hinders formation of misaminoacylated tRNAs, analysis of the mechanism of the editing reaction for class I enzyme leucyl-tRNA synthetase complexed with a misaminoacylated tRNALeu by initio hybrid quantum mechanical/molecular mechanical potentials in conjunction with molecular dynamics simulations, overview. Editing is a self-cleavage reaction of the tRNA and so it is the tRNA, and not the protein, that drives the reaction. The protein does, however, have an important stabilizing effect on some high-energy intermediates along the reaction path, which is more efficient than the ribozyme would be alone. This indicates that editing is achieved by a hybrid ribozyme/protein catalyst. The water molecule that acts as the nucleophile in the editing reaction is activated by a 3'-hydroxyl group at the 3'-end of tRNALeu and that the O2' atom of the leaving group of the substrate is capped by one of the water's hydrogen atoms
physiological function
-
aminoacyl-tRNA synthetases have evolved editing mechanisms to hydrolyze misactivated amino acids (pre-transfer editing) or misacylated tRNAs (post-transfer editing). Class Ia leucyl-tRNA synthetase may misactivate various natural and non-protein amino acids and then mischarge tRNALeu. The fidelity of prokaryotic LeuRS depends on multiple editing pathways to clear the incorrect intermediates and products in every step of aminoacylation reaction. Post-transfer editing as a final checkpoint of the reaction is very important to prevent mis-incorporation in vitro
physiological function
-
the carboxy-terminal domain of human mitochondrial leucyl-tRNA synthetase can be used to correct mitochondrial dysfunctions caused by mitochondrial tRNA mutations like the phenotype of m.3243A>G MTTL1 mutant cybrids
physiological function
-
the enzyme is a leucine sensor for serine/threonine kinase TORC1 and interacts with Gtr1
physiological function
-
the enzyme is a leucine sensor for serine/threonine kinase TORC1 and interacts with Gtr2
physiological function
-
the enzyme naturally produces mischarged tRNALeu
physiological function
-
the enzyme plays a critical role in amino acid-induced mammalian target of rapamycin C1 activation by sensing intracellular leucine concentration and initiating molecular events leading to mammalian target of rapamycin C1 activation. The enzyme directly binds to Rag GTPase, the mediator of amino acid signaling to mTORC1, in an amino acid-dependent manner and functions as a GTPase-activating protein for Rag GTPase to activate mammalian target of rapamycin C1
metabolism
-
isoform LeuRS1 generates Leu-tRNALeu for protein biosynthesis and exhibits obvious post-transfer editing activity to prevent generation of mischarged tRNALeu
-
additional information
-
activating role of C-terminal domain in the reactions of aminoacylation and editing, and its contribution to interaction with tRNALeu, overview. The C-terminal domain does is not critical for the manifestation of specificity of the enzyme of homologous RNAs, but is required for to enhance the rate of catalysis in aminoacylation and editing reaction
additional information
-
human leucyl-tRNA synthetase and mitochondrial protein elongation factor EF-Tu show suppressing cross-activity on different tRNA mutants in humans and Saccharomyces cerevisiae, mechanism and specificity of suppression, overview. Suppressive activities of wild-type and mutant enzymes, overview
additional information
-
leucine-specific domain 1, LSD1, is dispensable for post-transfer editing
additional information
-
the CP1, i.e. connective peptide 1, domain of LeuRS contains the editing active site ESI, eukarya-specific insertion 1, Thr341 serves as a specificity discriminator. Arg338 is crucial for tRNALeu charging and the Asp440 is crucial for leucine activation and aminoacylation. The post-transfer editing required the C-terminal domain, Arg338 and Asp440 of GlLeuRS
SUBSTRATE
PRODUCT                      
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
2'-dATP + L-leucine + tRNALeu
2'-dAMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
3'-dATP + L-leucine + tRNALeu
3'-dAMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
3'-dATP + L-leucine + tRNALeu
3'-dAMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
3'-dATP + L-leucine + tRNALeu
3'-dAMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Escherichia coli MRE 600
-
-
-
-
-
8-azaadenosine 5'-triphosphate + L-leucine + tRNALeu
8-azaadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
8-bromoadenosine 5'-triphosphate + L-leucine + tRNALeu
8-bromoadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
8-bromoadenosine 5'-triphosphate + L-leucine + tRNALeu
8-bromoadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
8-methylaminoadenosine 5'-triphosphate + L-leucine + tRNALeu
8-methylaminoadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
Adenosine 5'-O-(3-thio)triphosphate + L-leucine + tRNALeu
adenosine 5'-monophosphate + thiodiphosphate + L-leucyl-tRNALeu
show the reaction diagram
Escherichia coli, Escherichia coli MRE 600
-
-
-
-
-
Adenylyl beta,gamma-imido diphosphonate + L-leucine + tRNALeu
Adenylic acid + imido-diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
Adenylyl beta,gamma-imido diphosphonate + L-leucine + tRNALeu
Adenylic acid + imido-diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Escherichia coli MRE 600
-
-
-
-
-
AMP + diphosphate + Ile-tRNALeu
ATP + L-isoleucine + tRNALeu
show the reaction diagram
-
-
-
-
-
AMP + diphosphate + L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
ATP + L-leucine + Pyrococcus horikoshii tRNALeu(GAG)
show the reaction diagram
-
-
-
-
r
ATP + 2-butynylalanine + tRNALeu
AMP + diphosphate + 2-butynylalanyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + allylglycine + tRNALeu
AMP + diphosphate + allylglycyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + homoallylglycine + tRNALeu
AMP + diphosphate + homoallylglycyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + homopropargylglycine + tRNALeu
AMP + diphosphate + homopropargylglycyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + L-didehydroleucine + tRNALeu
AMP + diphosphate + didehydroleucyl-tRNALeu
show the reaction diagram
-
reaction is catalyzed by mutant T252Y, not by wild-type
-
-
?
ATP + L-isoleucine + 2'-deoxaadenosine-tRNALeu
AMP + ?
show the reaction diagram
-
2'-deoxyadenosine-tRNA clearly stimulates AMP production in the presence of isoleucine, but not the cognate leucine substrate
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
Q9P2J5
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
P26637
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
Q72GM3
mutant D345A, not the wild-type which performs only the misacetylation with isoleucine, but eliminates the incorrect isoleucyl-AMP
-
r
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
mutant D345A, not the wild-type which performs only the misacetylation with isoleucine, but eliminates the incorrect isoleucyl-AMP
-
r
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
mutant D419A, not the wild-type, which performs only the misacetylation with isoleucine, but eliminates the incorrect isoleucyl-AMP
-
r
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
wild-type and CP1 domain mutant enzyme, the mischarged product can be edited by the wild-type enzyme, but not by a recombinant isolated CP1 domain
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
activity with mutant enzyme D332A, no activity with wild-type full-length enzyme
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
activity with mutant enzymes T252E and T252D, no activity with wild-type enzyme and with mutant enzyme T252G
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
the ratio of turnover number to KM-value for L-leucine is 1600fold higher than the ratio observed for L-isoleucine
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
the ratio of turnover number to KM-value for L-leucine is 3000fold higher than the ratio observed for L-isoleucine
-
-
?
ATP + L-leucine + Natrialba magadii tRNALeu(CAA)
AMP + diphosphate + L-leucyl-Natrialba magadii tRNALeu(CAA)
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + Natrialba magadii tRNALeu(GAG)
AMP + diphosphate + L-leucyl-Natrialba magadii tRNALeu(GAG)
show the reaction diagram
-
highest activity
-
-
?
ATP + L-leucine + Pyrococcus horikoshii tRNALeu(GAG)
AMP + diphosphate + L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
show the reaction diagram
-
100% activity
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q72GM3
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q9P2J5
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q15031
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
P26637
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
r
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
acceptor activity with Tritrichomonas augusta tRNA is 8-fold higher than with yeast tRNA and 25-fold higher than with E. coli tRNA
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
chloroplastic enzyme: high specificity towards tRNAs, in contrast the cytoplasmic enzyme recognizes tRNAs from the bleached mutant and from yeast, but also some tRNALeu isoacceptors from E. coli
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
apart from the homologous substrate the enzyme is able to aminoacylate pure E. coli tRNALeu
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
cytoplasmic enzyme shows less strict specificity towards tRNA than the chloroplastic enzyme
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
enzyme activity only appears when both gene products, of leuS and leuS', coexist
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
mutant T252A edits correctly charged Leu-tRNALeu
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the connecting peptide CP1 domain is crucial for the editing function
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the cross-species-specific recognition occurs at the alpha-subunit, tRNALeu substrates from Escherichia coli and Aquifex aeolicus
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q72GM3
the editing active site hydrolytically cleaves the misactivated aminoacyl-adenylate, called pre-transfer editing, or the mischarged tRNA, called post-transfer editing
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the peptide bond between Glu292 and Ala293 in the large connecting polypeptide CP1 is essential for activity
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
tRNALeu from Aquifex aeolicus and Escherichia coli, native and recombinant wild-type, the recombinant isolated beta-subunit is inactive in catalysis
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q15031
tRNALeu substrate from Escherichia coli, 2-step reaction, the first step is reversible, while the second step is not
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
tRNALeu substrates from Escherichia coli and Aquifex aeolicus
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
two-step reaction, the beta-subunit alone is responsible for cognate tRNA recognition, enzyme activity requires both subunits
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
two-step reaction, the connecting peptide CP1 domain is crucial for the editing function
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
two-step reaction, the first step is reversible, the second is not, tRNA discrimination by a double-sieve mechansim
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the reaction catalyzed by the enzyme plays an important role in the transport of aminoacylated tRNAs from the nucleus to the cytoplasm
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
human cytosolic leucyl-tRNA synthetase is one component of a macromolecular aminoacyl-tRNA synthetase complex. The C-terminal peptide of hcLeuRS is critical for the interaction with hcArgRS and the interaction in the multi-tRNA synthetase complex
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
P07813
it is proposed that the enzyme uses a lock-and-key mechanism to recognize and discriminate the amino acids
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
two distinct domains of the beta subunit of Aquifex aeolicus leucyl-tRNA synthetase are involved in tRNA binding as revealed by a three-hybrid selection
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
a complex between prolyl-tRNA synthetase, ProRS, and LeuRS in Methanothermobacter thermautotrophicus enhances tRNAPro aminoacylation, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
possibly the yeast mitochondria have evolved to tolerate lower levels of fidelity in protein synthesis or have developed alternate mechanisms to enhance discrimination of leucine from non-cognate amino acids that can be misactivated by leucyl-tRNA synthetase
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
two functions of the enzyme in splicing and aminoacylation in vivo, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
a two step reaction, the C-terminal domain recognizes the long variable arm of tRNALeu for aminoacylation, and the so-called editing domain deacylates incorrectly formed Ile-tRNALeu, structural superposition of tRNAIle onto the LeuRS-tRNALeu complex indicated that Ile911, Lys912, and Glu913 of the LeuRS C-terminal domain clash with U20 of tRNAIle, which is bulged out as compared to the corresponding nucleotide of tRNALeu, mechanism for prevention of misediting, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
a two step reaction, the first of which is reversible, aminoacylation and editing by LeuRS require migration of the tRNA acceptor stem end between the canonical aminoacylation core and a separate domain called CP1 that is responsible for amino acid editing, post-transfer editing mechanism., overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
a two step reaction, the first of which is reversible, overview, the unique inserted leucine-specific domain of LeuRS is required for aminoacylation and not amino acid editing, the domain interacts with the tRNA during amino acid activation and/or tRNA aminoacylation, it might aid the dynamic translocation process that moves tRNA from the aminoacylation to the editing complex, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
a two-step reaction, the first of which, the amino acid activation step, is reversible, while the second aminoacylation step is not, the amino acid editing site for LeuRS resides within the homologous CP1 domain, some positions are idiosyncratic to LeuRS including a conserved arginine conferring amino acid substrate recognition, it complements other sites in the amino acid binding pocket of the editing active site of Escherichia coli LeuRS, including Thr252 and Val338, the latter is second to the first, which collectively fine-tune amino acid specificity to confer fidelity, editing mechanism, residues Arg249, Asp251, Thr252, Met336, and Val338 are involved, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
human mitochondrial LeuRS achieves high aminoacylation fidelity without a functional editing active site, representing a rare example of a class I aminoacyl-tRNA synthetase that does not proofread its products, K600 strongly impacts aminoacylation in two ways: it affects both amino acid discrimination and tRNA binding, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
L570 strongly impacts aminoacylation in two ways: it affects both amino acid discrimination and tRNA binding, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
LeuRS has a hydrolytic active site that resides in a discrete amino acid editing domain called CP1, LeuRS misactivates many non-leucine amino acids, including isoleucine, valine, methionine, and also structurally similar metabolic cellular intermediate, but the enzyme has an editing active site that is competent for post-transfer editing of mischarged tRNA
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
recombinant tRNALeu substrate, two peptides of eight and nine amino acid residues in the domain located in the alpha subunit are essential for the enzymes activity
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the editing domain called CP1 is required for hydrolyzing the incorrectly misaminoacylated noncognate amino acids Ile and Val, the beta-strands, which link the CP1 domain to the aminoacylation core of LeuRS, are required for editing of mischarged tRNALeu, hydrolytic activity is also enhanced by inclusion of short flexible peptides, called hinges, at the end of both LeuRS beta-strands, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the enzyme hydrolyzes mischarged tRNAs through a post-transfer editing mechanism, the enzyme from Aquifex aeolicus edits the complete set of aminoacylated tRNAs generated by the three enzymes, leucyl-, isoleucyl-, and valyl-tRNA synthetases: Ile-tRNAIle, Val-tRNAIle, ValtRNAVal, Thr-tRNAVal, and Ile-tRNALeu, model of a primitive editing system containing a composite minihelix carrying the triple leucine, isoleucine, and valine identity mimicking the primitive tRNA precursor, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
recognition of tRNALeu by the leucyl-tRNA synthetase (LeuRS) is studied by RNA probing and mutagenesis. Results show that the base A73, the core structure of tRNA formed by the tertiary interactions U8-A14, G18-U55 and G19-C56, and the orientation of the variable arm are critical elements for tRNALeu aminoacylation. Although dispensable for aminoacylation, the anticodon arm carries discrete editing determinants that are required for stabilizing the conformation of the post-transfer editing state and for promoting translocation of the tRNA acceptor arm from the synthetic to the editing site
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
tRNAs from Homo sapiens and Giardia lamblia
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Escherichia coli MRE 600
-
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Cyberlindnera jadinii Torulopsis
-
apart from the homologous substrate the enzyme is able to aminoacylate pure E. coli tRNALeu
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
?
show the reaction diagram
-
esterifies L-leucine to the cognate tRNA in the initial step of protein biosynthesis
-
-
-
ATP + L-leucine + tRNALeu
?
show the reaction diagram
-
mitochondrial enzyme is involved in protein synthesis and mRNA splicing
-
-
-
ATP + L-leucine + tRNALeu(GAG)
AMP + diphosphate + L-leucyl-tRNALeu(GAG)
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu(GAG)
AMP + diphosphate + L-leucyl-tRNALeu(GAG)
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu(UAA)
AMP + diphosphate + L-leucyl-tRNALeu(UAA)
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu(UAG)
AMP + diphosphate + L-leucyl-tRNALeu(UAG)
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu(UUR)
AMP + diphosphate + L-leucyl-tRNALeu(UUR)
show the reaction diagram
-
leucyl-tRNA synthetase contacts tRNALeu(UUR) in the amino acid acid acceptor stem, the anticodon stem, and the D-loop
-
-
?
ATP + L-leucine + tRNALeuA35G
AMP + diphosphate + L-leucyl-tRNALeuA35G
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeuA73
AMP + diphosphate + L-leucyl-tRNALeuA73
show the reaction diagram
-
class II tRNALeu, recognition requires the discriminator base A73 and the long variable arm of appropriate stem length, especially the specific loop sequence A47CG47D and U47H at the base of the helix
-
?
ATP + L-leucine + tRNALeuA73G
AMP + diphosphate + L-leucyl-tRNALeuA73G
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeuU73
AMP + diphosphate + L-leucyl-tRNALeuU73
show the reaction diagram
-
class II tRNALeu isoacceptor, 17fold lower activity compared to tRNALeuA73
-
?
ATP + L-leucine + tRNASer mutant
AMP + diphosphate + L-leucyl-tRNASer mutant
show the reaction diagram
-
transplantation of both the discriminator base and the variable arm of tRNALeu are not sufficient to introduce leucylation activity to tRNASer, but additional insertion of additional a nucleotide into the D-loop, which is not involved in the direct interaction with the enzyme, converts tRNASer to an efficient leucine acceptor
-
?
ATP + L-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
show the reaction diagram
-
mutant D345A, not the wild-type enzyme
-
r
ATP + L-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
show the reaction diagram
-
mutant D419A, not the wild-type enzyme
-
r
ATP + L-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
show the reaction diagram
-
wild-type and CP1 domain mutant enzyme, the mischarged product can be edited by the wild-type enzyme, but not by a recombinant isolated CP1 domain
-
?
ATP + L-norisoleucine + tRNALeu
AMP + diphosphate + L-norisoleucyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
show the reaction diagram
P26637
-
-
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + L-oxonorvaline + tRNALeu
AMP + diphosphate + oxonorvalyl-tRNALeu
show the reaction diagram
-
reaction is catalyzed by mutant T252Y, not by wild-type
-
-
?
L-isoleucyl-tRNALeu + H2O
t-RNALeu + isoleucine
show the reaction diagram
-
-
editing activity
-
?
tubercidin 5'-triphosphate + L-leucine + tRNALeu
tubercidin 5'-phosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
tubercidin 5'-triphosphate + L-leucine + tRNALeu
tubercidin 5'-phosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
-
tubercidin 5'-triphosphate + L-leucine + tRNALeu
tubercidin 5'-phosphate + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Escherichia coli MRE 600
-
-
-
-
-
L-isoleucyl-tRNALeu + H2O
t-RNALeu + L-isoleucine
show the reaction diagram
Q9P2J5
-
editing activity
-
?
additional information
?
-
-
leucine-dependent ATP-diphosphate exchange, leucine + ATP + enzyme/Ile-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
leucine-dependent ATP-diphosphate exchange, leucine + ATP + enzyme/Ile-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
leucine-dependent ATP-diphosphate exchange, leucine + ATP + enzyme/Ile-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
leucine-dependent ATP-diphosphate exchange, leucine + ATP + enzyme/Ile-AMP-enzyme + diphosphate
-
-
-
additional information
?
-
-
no substrates: dATP, GTP, dGTP
-
-
-
additional information
?
-
-
activity with mitochondrial tRNA mutants associated with some human mitochondrion-related neuromuscular disorders
-
?
additional information
?
-
Q15031
enzyme also performs the ATP-diphosphate exchange reaction
-
?
additional information
?
-
-
proteolytically derived 34 kDa peptide fragment has lost most of its aminoacylation activity, but retains the ATP-dihosphate exchnage activity, the enzyme also performs the ATP-diphosphate exchange reaction
-
?
additional information
?
-
-
substrate specificty with diverse class II tRNALeu isoacceptors and mutants, overview, no activity with tRNALeuG73 and C73, no activity with tRNALeu, tRNASer and tRNATyr from Escherichia coli and Saccharomyces cerevisiae, differences in the tertiary structure of tRNALeu and tRNASer play a key role for inactivity and therefore elimination of native tRNASer as leucine acceptor
-
?
additional information
?
-
-
the enzyme also performs the ATP-diphosphate exchange reaction
-
?
additional information
?
-
-
the enzyme also performs the ATP-diphosphate exchange reaction
-
?
additional information
?
-
-
the enzyme also performs the ATP-diphosphate exchange reaction, the enzyme has an editing function to correct misaminoacylation of tRNALeu by isoleucine and methionine, T252 is involved
-
?
additional information
?
-
-
the enzymes forms also perform the reversible ATP-diphosphate exchange reaction, which corresponds to the first reaction step
-
?
additional information
?
-
-
aminoacyl-tRNA is channeled in vivo by probably direct transfer to elongation factor I
-
?
additional information
?
-
-
fidelity of translation is dependent on the specificity of the aminoacyl-tRNA synthetases
-
?
additional information
?
-
-
aminoacylation of minihelices is strongly dependent on the presence of the A73 identity nucleotide and greatly stimulated by destabilization of the first base pair. Addition of RNA helices that mimic the anticodon domain stimulates minihelixLeu charging by alphabeta-LeuRS indicating possible domain-domain communication. MinihelixLeu cannot be misaminoacylated, perhaps because of the tRNA-independent pretransfer editing activity of alphabeta-LeuRS
-
-
-
additional information
?
-
-
isolated editing domain of leucyl-tRNA synthetase from the deep-rooted bacterium Aquifex aeolicus catalyzes the hydrolytic editing of both mischarged tRNALeu and minihelixLeu
-
-
-
additional information
?
-
-
Thr247 and Thr248 are two key residues in the Escherichia coli LeuRS editing active site and appear to collaborate in the hydrolytic cleavage mechanism
-
-
-
additional information
?
-
-
wild-type, full-length enzyme deacylates the pre-formed Ile-tRNALeu
-
-
-
additional information
?
-
-
isolated LeuRS CP1 domain requires idiosyncratic adaptations to confer editing activity independent of the full-length enzyme, overview
-
-
-
additional information
?
-
-
the LeuRS CP1 domain can also support group I intron RNA splicing in the yeast mitochondria, overview, the RDW peptide, a highly conserved peptide within an RDW-containing motif, is important for enzyme interactions, the RDW peptide is dynamic and forms unique sets of interactions with the aminoacylation and editing complexes, overview
-
-
-
additional information
?
-
-
residues Y515 and Y520 outside the editing active site of CP1 domain of Giardia lamblia LeuRS are crucial for post-transfer editing by influencing the binding affinity with mischarged tRNALeu
-
-
-
additional information
?
-
-
LeuRS misactivates several non-cognate amino acids, e.g. Ile and Met as well as the non-standard amino acids norvaline and alpha-amino butyrate. It uses mainly pre-transfer editing to edit alpha-amino butyrate and a tRNA-dependent mechanism to edit norvaline, although both amino acids can be charged to tRNALeu, overview. Separation of the norvaline-editing pathways
-
-
-
additional information
?
-
-
measurement of ATP-PPi exchange activity by wild-type and mutant enzymes
-
-
-
additional information
?
-
-
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
-
-
-
additional information
?
-
-
the enzyme maintains weak pre-transfer editing activities
-
-
-
additional information
?
-
-
the enzyme maintains weak pretransfer editing activities
-
-
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
LITERATURE
(Substrate)
COMMENTARY
(Product)
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + 2-butynylalanine + tRNALeu
AMP + diphosphate + 2-butynylalanyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + allylglycine + tRNALeu
AMP + diphosphate + allylglycyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + homoallylglycine + tRNALeu
AMP + diphosphate + homoallylglycyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + homopropargylglycine + tRNALeu
AMP + diphosphate + homopropargylglycyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q72GM3
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
Q15031
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
the reaction catalyzed by the enzyme plays an important role in the transport of aminoacylated tRNAs from the nucleus to the cytoplasm
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
human cytosolic leucyl-tRNA synthetase is one component of a macromolecular aminoacyl-tRNA synthetase complex. The C-terminal peptide of hcLeuRS is critical for the interaction with hcArgRS and the interaction in the multi-tRNA synthetase complex
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
a complex between prolyl-tRNA synthetase, ProRS, and LeuRS in Methanothermobacter thermautotrophicus enhances tRNAPro aminoacylation, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
possibly the yeast mitochondria have evolved to tolerate lower levels of fidelity in protein synthesis or have developed alternate mechanisms to enhance discrimination of leucine from non-cognate amino acids that can be misactivated by leucyl-tRNA synthetase
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
two functions of the enzyme in splicing and aminoacylation in vivo, overview
-
-
?
ATP + L-leucine + tRNALeu
?
show the reaction diagram
-
esterifies L-leucine to the cognate tRNA in the initial step of protein biosynthesis
-
-
-
ATP + L-leucine + tRNALeu
?
show the reaction diagram
-
mitochondrial enzyme is involved in protein synthesis and mRNA splicing
-
-
-
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
-
?
ATP + L-norisoleucine + tRNALeu
AMP + diphosphate + L-norisoleucyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
show the reaction diagram
-
aminoacylation by mutant T252Y
-
?
additional information
?
-
-
aminoacyl-tRNA is channeled in vivo by probably direct transfer to elongation factor I
-
?
additional information
?
-
-
fidelity of translation is dependent on the specificity of the aminoacyl-tRNA synthetases
-
?
additional information
?
-
-
LeuRS misactivates several non-cognate amino acids, e.g. Ile and Met as well as the non-standard amino acids norvaline and alpha-amino butyrate. It uses mainly pre-transfer editing to edit alpha-amino butyrate and a tRNA-dependent mechanism to edit norvaline, although both amino acids can be charged to tRNALeu, overview. Separation of the norvaline-editing pathways
-
-
-
additional information
?
-
-
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
-
-
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ATP
-
optimal concentration is 4 mM for the wild-type and mutant A293R, for the mutants A293Y, A293G, and A293I it is 2 mM, for the mutants A293D, and As93F it is 1 mM
ATP
-
optimal concentration for the wild-type enzyme is 4 mM, for the mutants 2 mM
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Al3+
-
accelerates reaction
Ba2+
-
Mg2+, Mn2+, Ca2+ and Ba2+ similarly effective, optimal concentration: 0.1 mM
Ca2+
-
Mg2+, Mn2+, Ca2+ and Ba2+ similarly effective, optimal concentration: 0.1 mM
Ca2+
-
75% of the maximal activity obtained with Mg2+; can partially replace Mg2+ in activation (in chloroplastic enzyme, not in cytoplasmic enzyme)
Ca2+
-
can partially replace Mg2+ in activation (in chloroplastic enzyme, not in cytoplasmic enzyme)
Co2+
-
40% of the maximal activity obtained with Mg2+; can partially replace Mg2+ in activation (in chloroplastic enzyme, not in cytoplasmic enzyme
Co2+
-
can partially replace Mg2+ in activation (in chloroplastic enzyme, not in cytoplasmic enzyme
Fe2+
-
accelerates reaction
Fe3+
-
accelerates reaction
K+
-
optimal concentration: 80 mM KCl
K+
-
5.0 mM, mitochondrial enzyme, 2.5 mM, cytoplasmic enzyme
K+
-
slight stimulation by 50 mM KCl, progressive inhibition with higher concentrations
K+
-
30 mM KCl enhance activity of cytoplasmic enzyme, no effect on chloroplastic enzyme, concentrations higher than 30 mM inhibit cytoplasmic enzyme more strongly than chloroplastic enzyme
K+
-
the enzyme activity requires high KCl concentration with highest activity in the presence of 3.75 M KCl
Mg2+
-
activates; optimal concentration: 14 mM Mg-acetate
Mg2+
-
Mg2+, Mn2+, Ca2+ and Ba2+ similarly effective, optimal concentration: 0.1 mM
Mg2+
-
activates; optimal concentration: 4 mM (with an ATP concentration of 2 mM)
Mg2+
-
optimal concentration: 2.5 mM (mitochondrial enzyme), 10 mM (cytoplasmic enzyme)
Mg2+
-
activates; optimal Mg2+/ATP ratio: 1.2
Mg2+
-
optimal Mg2+/ATP ratio: 1.5
Mg2+
-
optimal concentration is 12 mM
Mg2+
-
-
Mg2+
-
-
Mg2+
-
required
Mg2+
-
required
Mg2+
-
the optimum Mg2+:ATP ratio for aminoacylation is 3:1
Mn2+
-
Mg2+, Mn2+, Ca2+ and Ba2+ similarly effective, optimal concentration: 0.1 mM
Mn2+
-
75% of the maximal activity obtained with Mg2+; can partially replace Mg2+ in activation (in chloroplastic enzyme, not in cytoplasmic enzyme)
Mn2+
-
can partially replace Mg2+ in activation (in chloroplastic enzyme, not in cytoplasmic enzyme)
Sn2+
-
accelerates reaction
Sn4+
-
accelerates reaction
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
(2E)-3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)-1-phenylprop-2-en-1-one
-
-
(2E)-3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-7-yl)-1-phenylprop-2-en-1-one
-
-
(E)-[3-(1,3-dihydro-1-hydroxy-2,1-benzoxaborol-7-yl)]acrylic acid ethyl ester
-
-
1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl cyclohexylcarbamate
-
-
1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl phenylcarbamate
-
-
1-hydroxy-1,3-dihydro-2,1-benzoxaborole-7-carbaldehyde
-
-
1-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-4-methylpentan-2-one
-
-
1-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]butan-2-one
-
-
1-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]pentan-2-one
-
-
2'-dATP
-
-
2,1-benzoxaborol-1(3H)-ol
-
-
2,1-benzoxaborole-1,6(3H)-diol
-
-
2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]pentan-3-one
-
-
2-[3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]propyl]-1H-isoindole-1,3(2H)-dione
-
-
3'-Amino-3'-deoxy adenosine 5'-triphosphate
-
-
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-3-methylbutan-2-one
-
-
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-4-methylpentan-2-one
-
-
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]butan-2-one
-
-
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]heptan-4-one
-
-
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]hexan-2-one
-
-
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]pentan-2-one
-
-
5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole
-
i.e. AN2690, 0.1 mM, 5fold decrease in aminoacylation activity
5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole
-
i.e. AN2690, the editing active site is the proven target for the broad-spectrum drug. But the post-transfer editing by LeuRS is resistant to the broad-spectrum drug AN2690, AN2690 resistance and its possible mechanism, overview
5-fluoro-2,1-benzoxaborol-1(3H)-ol
-
AN-2690, antibiotic which specifically targets the editing active site of LeuRS
5-fluoro-2,1-benzoxaborol-1(3H)-ol
P26637
AN-2690, antibiotic which specifically targets the editing active site of LeuRS
6-(2,2-dimethoxyethoxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(2-methoxyethoxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(3-hydroxypropyl)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(benzyloxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(cyclohexylmethoxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(propan-2-yloxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(pyridin-2-ylmethoxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-(quinolin-2-yloxy)-2,1-benzoxaborol-1(3H)-ol
-
-
6-butoxy-2,1-benzoxaborol-1(3H)-ol
-
-
6-dimethylaminopurine riboside 5'-triphosphate
-
-
6-ethoxy-2,1-benzoxaborol-1(3H)-ol
-
-
6-mercaptopurine riboside 5'-triphosphate
-
-
6-methylaminopurine riboside 5'-triphosphate
-
-
6-propoxy-2,1-benzoxaborol-1(3H)-ol
-
-
6-[(2-fluorobenzyl)oxy]-2,1-benzoxaborol-1(3H)-ol
-
-
6-[(3-hydroxypentan-2-yl)oxy]-2,1-benzoxaborol-1(3H)-ol
-
-
7-(3-hydroxypropyl)-2,1-benzoxaborol-1(3H)-ol
-
-
Adenine arabinoside 5'-triphosphate
-
-
adenosine
-
-
adenyl(alpha,beta-methylene)triphosphonate
-
-
adenylyl(beta,gamma-imido)triphosphonate
-
-
adenylyl(beta,gamma-methylene)diphosphonate
-
-
Al3+
-
in vitro the enzyme is inhibited by 40% at 0.04 mM, Al3+ inhibits the enzyme in vivo and in vitro, quantitative analysis, in vivo acceptor activity of tRNALeu is decreased by 23% thereby the leucyl-tRNA synthetase activity is increased by 20%, overview
ATP
-
at high concentration, the A293 mutants are ore sensitive
ATP
-
at high concentration, the mutants are more sensisitve than the wild-type enzyme
Cd2+
-
in presence of 0.1 mM Mg2+
diphosphate
-
-
ethyl (2E)-3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)prop-2-enoate
-
-
ethyl 2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-2-methylpropanoate
-
-
ethyl 2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]butanoate
-
-
ethyl 2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]propanoate
-
-
ethyl 3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)propanoate
-
-
ethyl 3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-7-yl)propanoate
-
-
ethyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy](phenyl)acetate
-
-
ethyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetate
-
-
N-ethyl-2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetamide
-
-
N-tert-butyl-2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetamide
-
-
NaCl
-
no activity of isoform LeuRS1 is detected in NaCl solutions
Nalidixic acid
-
-
norvaline
-
-
novobiocin
-
-
O-[N-(L-norvalyl)sulfamoyl]adenosine
Q72GM3
analogue to the reaction intermediate, non-hydrolyzable
Oxolinic acid
-
-
p-chloromercuribenzoate
-
-
p-chloromercuribenzoate
-
-
Purine riboside 5'-triphosphate
-
-
tert-butyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetate
-
-
tert-butyl [2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]ethyl]carbamate
-
-
Zn2+
-
in presence of 0.1 mM Mg2+
[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetaldehyde
-
-
[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetic acid
-
-
methyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetate
-
-
additional information
Q15031
inhibition by high levels of mono- and divalent cations
-
additional information
-
aminoacylation and editing reaction are resistant to inactivation by compound AN2690
-
additional information
-
development of a GlLeuRS-specific inhibitor for the treatment of giardiasis
-
additional information
-
enzyme drug inhibitor design and development based on the benzoxaborole structure, inhibitory potencies and effectiveness a anti-trypanosomal drugs, ligand, i.e. benzoxaborole-AMP, docking in the LeuRS homology model, overview
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
Al3+
-
in vivo acceptor activity of tRNALeu is decreased by 23% thereby the leucyl-tRNA synthetase activity is increased by 20%, overview
cadmium
-
-
putrescine
-
optimal concentration: 5.5 mM
spermidine
-
optimal concentration: 3 mM
spermidine
-
in absence of Mg2+ spermine allows a very low level of cytoplasmic enzyme
tRNALeu
-
presence of tRNALeu robustly stimulates editing activity
additional information
-
a complex between prolyl-tRNA synthetase, ProRS, and LeuRS in Methanothermobacter thermautotrophicus enhances tRNAPro aminoacylation, overview
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.045
3'-dATP
-
-
0.16
8-azaadenosine 5'-triphosphate
-
-
1
8-bromoadenosine 5'-triphosphate
-
-
1
8-bromoadenosine 5'-triphosphate
-
3'-dATP, tubercidin triphosphate
0.07
8-Methylaminoadenosine 5'-triphosphate
-
-
0.055
adenosine 5'-O-(3-thiotriphosphate)
-
-
0.08
ATP
-
mutant E292F, pH 7.8, 37C
0.09
ATP
Q15031
recombinant mitochondrial isozyme, pH 7.6, 37C
0.1
ATP
-
mutants E292Q, E292D, and E292A, pH 7.8, 37C
0.11
ATP
-
aminoacylation reaction, pH 7.8, 60C
0.11
ATP
-
mutants E292D and E292K, pH 7.8, 37C
0.112
ATP
-
65C, wild-type enzyme
0.1123
ATP
-
pH 6.8, 65C, recombinant wild-type enzyme
0.113
ATP
-
65C, recombinant His6-tagged enzyme
0.13
ATP
-
-
0.18
ATP
-
-
0.22
ATP
-
37C, pH 7.8, mutant enzyme T252E
0.228
ATP
-
recombinant enzyme complex, 65C
0.23
ATP
-
37C, pH 7.8, mutant enzyme T25D
0.24
ATP
-
37C, pH 7.8, native enzyme
0.25
ATP
-
aminoacylation reaction, mutant enzyme, pH 7.8, 37C
0.26
ATP
-
wild-type enzyme, pH 7.8, 37C
0.26
ATP
-
native enzyme
0.28
ATP
-
aminoacylation reaction, wild-type enzyme, pH 7.8, 37C
0.28
ATP
-
recombinant enzyme
0.296
ATP
-
pH 7.5, 65C, mutant R106A
0.323
ATP
-
pH 7.5, 65C, mutant R97A
0.33
ATP
-
mutant lacking residues Q281 to D294, 45C
0.351
ATP
-
pH 7.5, 65C, mutant V108A
0.36
ATP
-
ATP-diphosphate exchange reaction, pH 7.8, 37C
0.362
ATP
-
pH 7.5, 65C, mutant D98A
0.366
ATP
-
pH 7.5, 65C, mutant K100A/Y109A
0.37
ATP
-
pH 7.5, 65C, mutant N96A
0.373
ATP
-
pH 7.5, 65C, mutant E114A
0.38
ATP
-
ATP-diphosphate exchange reaction, pH 7.8, 60C
0.383
ATP
-
pH 7.5, 65C, mutant T101A
0.472
ATP
-
pH 7.5, 65C, mutant W103A
0.531
ATP
-
pH 7.5, 65C, mutant K100A
0.537
ATP
-
pH 7.5, 65C, mutant Y105A
0.547
ATP
-
pH 7.5, 65C, mutant F119A
0.55
ATP
-
aminoacylation reaction, pH 7.8, 37C
0.551
ATP
-
pH 7.5, 65C, mutant D121A
0.578
ATP
-
pH 7.5, 65C, wild-type enzyme
0.584
ATP
-
pH 7.5, 65C, mutant K100A/Y105A
0.59 - 1
ATP
-
pH 7.5, 65C, mutant Y109A
0.653
ATP
-
pH 8.2, 45C, mutant W155A
0.675
ATP
-
pH 8.2, 45C, mutant Q154A
0.683
ATP
-
pH 8.2, 45C, mutant K170A
0.687
ATP
-
pH 8.2, 45C, mutant K166A
0.688
ATP
-
wild-type, 45C
0.698
ATP
-
pH 8.2, 45C, mutant K148A
0.711
ATP
-
pH 8.2, 45C, mutant S153A
0.725
ATP
-
37C, pH 7.6, leucylation, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.728
ATP
-
pH 8.2, 45C, mutant K142A
0.772
ATP
-
pH 8.2, 45C, mutant K141A
0.773
ATP
-
37C, pH 7.6, leucylation, full-length enzyme
0.796
ATP
-
pH 7.5, 65C, mutant I104A
0.812
ATP
-
pH 8.2, 45C, mutant K139A
0.822
ATP
-
pH 8.2, 45C, mutant K144A
0.834
ATP
-
pH 8.2, 45C, mutant K152A
0.837
ATP
-
pH 7.5, 65C, mutant E113A
0.99
ATP
-
recombinant mitochondrial isozyme mutant, 37C
1.025
ATP
-
pH 8.2, 45C, mutant D173A
1.157
ATP
-
pH 7.5, 65C, mutant T118A
1.169
ATP
-
pH 8.2, 45C, mutant E165A
1.308
ATP
-
37C, pH 7.6, ATP-diphosphate exchange, full-length enzyme
1.349
ATP
-
37C, pH 7.6, ATP-diphosphate exchange, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
2.129
ATP
-
pH 8.2, 45C, mutant E167A
2.177
ATP
-
pH 7.5, 65C, mutant I115A
0.002
Ile-tRNALeu
-
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
0.0021
Ile-tRNALeu
-
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
0.0024
Ile-tRNALeu
-
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
0.0025
Ile-tRNALeu
-
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
1.034
L-didehydroleucine
-
mutant T252Y
0.25
L-isoleucine
-
pH 7.5, 37C
0.698
L-isoleucine
-
37C
2.04
L-isoleucine
Q9P2J5
wild-type, pH 7.6, 30C
2.8
L-isoleucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37C
3.3
L-isoleucine
Q9P2J5
mutant D399A, pH 7.6, 30C
3.5
L-isoleucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37C
8.713
L-isoleucine
P26637
-
14
L-isoleucine
-
pH 7.5, 37C
0.0092
L-isoleucyl-tRNALeu
-
wild-type, pH 7.5, 37C
0.0122
L-isoleucyl-tRNALeu
-
mutant Y515A, pH 7.5, 37C
0.0147
L-isoleucyl-tRNALeu
-
mutant Y520H, pH 7.5, 37C
0.0173
L-isoleucyl-tRNALeu
-
mutant Y520A, pH 7.5, 37C
0.008
L-Leu
-
-
0.014
L-Leu
-
-
0.02
L-Leu
-
-
0.02
L-Leu
-
-
0.05
L-Leu
-
adenylyl (beta,gamma-imido)diphosphonate
0.05
L-Leu
-
-
0.0011
L-leucine
-
-
0.0013
L-leucine
-
ATP-diphosphate exchange reaction, pH 7.8, 37C
0.0015
L-leucine
-
wild-type enzyme, aminoacylation
0.0016
L-leucine
-
ATP-diphosphate exchange reaction, pH 7.8, 60C
0.0016
L-leucine
-
mutant enzyme T252V, aminoacylation
0.002
L-leucine
-
mutant enzyme T252S, aminoacylation
0.0024
L-leucine
-
pH 8.2, 45C, mutant K152A
0.0028
L-leucine
-
37C
0.0035
L-leucine
-
pH 8.2, 45C, mutant Q154A
0.0036
L-leucine
-
mutant enzyme T252A, aminoacylation
0.0041
L-leucine
P26637
-
0.00537
L-leucine
-
pH 7.5, 65C, mutant N96A
0.0054
L-leucine
-
mutant Y515A, pH 8.2, 45C; mutant Y515E, pH 8.2, 45C; mutant Y520H, pH 8.2, 45C
0.00555
L-leucine
-
pH 7.5, 65C, mutant Y105A
0.00559
L-leucine
-
pH 7.5, 65C, mutant I115A
0.00569
L-leucine
-
pH 7.5, 65C, mutant I104A
0.0057
L-leucine
-
pH 7.5, 65C, mutant K100A/Y109A
0.0058
L-leucine
-
37C, pH 7.6, leucylation, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.00581
L-leucine
-
pH 7.5, 65C, mutant V108A
0.0059
L-leucine
-
mutant lacking residues Q281 to D294, 45C
0.00592
L-leucine
-
pH 7.5, 65C, mutant D98A; pH 7.5, 65C, wild-type enzyme
0.006
L-leucine
-
aminoacylation reaction, pH 7.8, 60C
0.006
L-leucine
-
pH 6.8, 65C, recombinant wild-type enzyme
0.006
L-leucine
-
mutant Y520A, pH 8.2, 45C
0.0061
L-leucine
-
wild-type, pH 8.2, 45C
0.00618
L-leucine
-
pH 7.5, 65C, mutant E113A
0.00623
L-leucine
-
pH 7.5, 65C, mutant E114A
0.0063
L-leucine
-
pH 7.5, 65C, mutant K100A/Y105A
0.00636
L-leucine
-
pH 7.5, 65C, mutant T118A
0.0064
L-leucine
-
aminoacylation reaction, pH 7.8, 37C
0.0064
L-leucine
-
65C, recombinant His6-tagged enzyme; 65C, wild-type enzyme
0.00721
L-leucine
-
pH 7.5, 65C, mutant D121A
0.0075
L-leucine
-
37C, pH 7.6, leucylation, full-length enzyme
0.0077
L-leucine
-
mutant Y520E, pH 8.2, 45C
0.0077
L-leucine
-
mutant lacking residues S295 to L304, 45C
0.008
L-leucine
-
mutant Y515K, pH 8.2, 45C
0.0083
L-leucine
-
recombinant enzyme complex, 65C
0.00843
L-leucine
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0.00854
L-leucine
-
pH 7.5, 65C, mutant R106A
0.00871
L-leucine
-
pH 7.5, 65C, mutant W103A
0.00891
L-leucine
-
pH 7.5, 65C, mutant K100A
0.009
L-leucine
-
wild-type, 45C
0.00902
L-leucine
-
pH 7.5, 65C, mutant Y109A
0.00927
L-leucine
-
pH 7.5, 65C, mutant R97A
0.00984
L-leucine
-
pH 7.5, 65C, mutant T101A
0.01
L-leucine
-
recombinant mitochondrial isozyme mutant, 37C
0.0118
L-leucine
-
pH 7.5, 65C, mutant F119A
0.0119
L-leucine
-
pH 8.2, 45C, mutant W155A
0.012
L-leucine
-
mutant E292K, pH 7.8, 37C
0.0124
L-leucine
-
pH 8.2, 45C, mutant K148A
0.013
L-leucine
-
mutant E292S, pH 7.8, 37C
0.0132
L-leucine
-
pH 8.2, 45C, mutant K141A
0.014
L-leucine
-
mutant E292D, E292A, and E292F pH 7.8, 37C
0.0141
L-leucine
-
pH 8.2, 45C, mutant K170A
0.0145
L-leucine
-
pH 8.2, 45C, mutant K142A
0.0147
L-leucine
-
pH 8.2, 45C, mutant K166A
0.015
L-leucine
-
aminoacylation reaction, wild-type and mutant enzyme, pH 7.8, 37C
0.015
L-leucine
-
wild-type enzyme and mutant E292Q, pH 7.8, 37C
0.015
L-leucine
-
recombinant and native enzyme
0.015
L-leucine
-
pH 7.5, 37C
0.0151
L-leucine
-
pH 8.2, 45C, mutant K139A
0.0153
L-leucine
-
pH 8.2, 45C, mutant K144A
0.0157
L-leucine
-
pH 8.2, 45C, mutant E165A
0.0159
L-leucine
-
pH 8.2, 45C, mutant S153A
0.0162
L-leucine
-
pH 8.2, 45C, mutant E167A
0.0165
L-leucine
-
pH 8.2, 45C, mutant D173A
0.018
L-leucine
-
mutant T252Y
0.019
L-leucine
-
37C, pH 7.8, mutant enzyme T252E; 37C, pH 7.8, mutant enzyme T25D
0.02
L-leucine
-
37C, pH 7.8, native enzyme
0.021
L-leucine
-
recombinant mitochondrial isozyme mutant, 37C
0.024
L-leucine
Q9P2J5
wild-type, pH 7.6, 30C
0.039
L-leucine
-
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
0.045
L-leucine
Q15031
recombinant mitochondrial isozyme, pH 7.6, 37C
0.0456
L-leucine
-
pH 7.6, 37C, wild-type enzyme
0.05
L-leucine
-
pH 7.6, 37C, mutant D399A
0.052
L-leucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37C
0.064
L-leucine
-
37C, pH 7.6, ATP-diphosphate exchange, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.069
L-leucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37C
0.075
L-leucine
-
37C, pH 7.6, ATP-diphosphate exchange, full-length enzyme
0.13
L-leucine
-
pH 7.5, 37C
0.891
L-leucine
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0.9
L-leucine
Q9P2J5
mutant D399A, pH 7.6, 30C
0.01251
L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.01643
L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.983
L-methionine
-
37C
6.2
L-methionine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37C
7.5
L-methionine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37C
0.096
L-norvaline
-
37C
2.245
L-oxonorvaline
-
mutant T252Y
0.01101
Natrialba magadii tRNALeu(CAA)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.01313
Natrialba magadii tRNALeu(CAA)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.00335
Natrialba magadii tRNALeu(GAG)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.00773
Natrialba magadii tRNALeu(GAG)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.167
norvaline
P26637
-
0.0001
tRNALeu
-
-
0.00011
tRNALeu
-
37C, pH 7.8, mutant enzyme K238A
0.00014
tRNALeu
-
37C, pH 7.8, mutant enzyme G237D
0.00018
tRNALeu
-
37C, pH 7.8, mutant enzyme L283F
0.0002
tRNALeu
-
37C, pH 7.8, wilde-type enzyme
0.0002
tRNALeu
-
mutant R449K
0.00024
tRNALeu
-
37C, pH 7.8, mutant enzyme K160N
0.00025
tRNALeu
-
37C, pH 7.8, mutant enzyme Q234H
0.00029
tRNALeu
-
37C, pH 7.8, mutant enzyme N152A
0.0003
tRNALeu
-
-
0.00038
tRNALeu
-
37C, pH 7.8, mutant enzyme M159A
0.0004
tRNALeu
-
-
0.0005
tRNALeu
-
37C, pH 7.8, mutant enzyme R94A
0.0005
tRNALeu
-
mutant W445Y
0.00051
tRNALeu
-
37C, pH 7.8, mutant enzyme A156V
0.0006
tRNALeu
-
wild-type enzyme
0.00061
tRNALeu
-
37C, pH 7.8
0.0007
tRNALeu
-
wild-type enzyme
0.0007
tRNALeu
-
mutant R451K
0.00073
tRNALeu
-
pH 7.5, 37C, recombinant wild-type enzyme
0.00085
tRNALeu
-
substrate from E. coli
0.0009
tRNALeu
-
mutant V338A
0.0011
tRNALeu
-
mutant enzyme V910A, at pH 8.2 and 30C; wild type enzyme, at pH 8.2 and 30C
0.0012
tRNALeu
-
mutant E292K, pH 7.8, 37C
0.0012
tRNALeu
-
mutant enzyme L964A, at pH 8.2 and 30C
0.0013
tRNALeu
-
-
0.0013
tRNALeu
-
mutant enzyme Q915K, at pH 8.2 and 30C
0.0014
tRNALeu
-
37C, pH 7.6, tRNALeu from calf liver, leucylation, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.0014
tRNALeu
-
mutant enzyme V910W, at pH 8.2 and 30C
0.0015
tRNALeu
-
aminoacylation reaction, wild-type enzyme, pH 7.8, 37C
0.0016
tRNALeu
-
of Euglena gracilis
0.0016
tRNALeu
-
recombinant and native enzyme
0.00167
tRNALeu
-
full-length enzyme, pH 7.5, 37C
0.00179
tRNALeu
-
truncation mutant DELTA911-913, pH 7.5, 37C
0.0018
tRNALeu
-
mutant enzyme R921K, at pH 8.2 and 30C
0.0019
tRNALeu
-
37C, pH 7.6, tRNALeu from calf liver, leucylation, full-length enzyme
0.0019
tRNALeu
-
mutant enzyme Q915A, at pH 8.2 and 30C
0.002
tRNALeu
-
pH 7.5, 37C, recombinant LS-domain deletion mutant
0.0021
tRNALeu
-
mutant E292A, pH 7.8, 37C
0.0024
tRNALeu
-
aminoacylation reaction, mutant enzyme, pH 7.8, 37C
0.0024
tRNALeu
-
mutant E292S, pH 7.8, 37C
0.0025
tRNALeu
-
wild-type enzyme, pH 7.8, 37C
0.0025
tRNALeu
-
37C, pH 7.8, mutant enzyme T25D
0.0026
tRNALeu
-
unfractionated substrate of E. coli
0.0026
tRNALeu
-
37C, pH 7.8, native enzyme
0.0031
tRNALeu
-
mutant E292Q, pH 7.8, 37C
0.0033
tRNALeu
-
37C, pH 7.8, mutant enzyme T252E
0.0035
tRNALeu
-
37C, pH 7.8, mutant enzyme V286stop
0.0035
tRNALeu
-
pH 8.2, 45C, mutant K142A
0.0038
tRNALeu
-
pH 8.2, 45C, mutant Q154A
0.004
tRNALeu
-
mutants E292D and E292F, pH 7.8, 37C
0.004
tRNALeu
-
pH 8.2, 45C, mutant S153A
0.0049
tRNALeu
-
pH 8.2, 45C, mutant E167A
0.005
tRNALeu
-
pH 8.2, 45C, mutant K148A
0.0051
tRNALeu
-
pH 8.2, 45C, mutant K141A; pH 8.2, 45C, mutant K152A; pH 8.2, 45C, mutant K170A
0.0052
tRNALeu
-
pH 8.2, 45C, mutant K144A
0.0052
tRNALeu
-
mutant enzyme R921A, at pH 8.2 and 30C
0.0056
tRNALeu
-
pH 8.2, 45C, recombinant mutant T341A
0.0057
tRNALeu
-
pH 8.2, 45C, recombinant mutant D444A
0.0059
tRNALeu
-
pH 8.2, 45C, mutant D173A
0.0059
tRNALeu
-
pH 8.2, 45C, recombinant mutant T341R
0.0061
tRNALeu
-
wild-type, 45C
0.0071
tRNALeu
-
pH 8.2, 45C, mutant W155A
0.008
tRNALeu
-
pH 8.2, 45C, recombinant mutant R338A
0.0081
tRNALeu
-
pH 8.2, 45C, mutant K166A
0.0082
tRNALeu
-
mutant lacking residues S295 to L304, 45C
0.0083
tRNALeu
-
pH 8.2, 45C, mutant E165A
0.0083
tRNALeu
-
mutant enzyme L949A, at pH 8.2 and 30C
0.0092
tRNALeu
-
mutant enzyme V910P, at pH 8.2 and 30C
0.0093
tRNALeu
-
pH 8.2, 45C, recombinant mutant DELTAESI/DELTAHsESI
0.0095
tRNALeu
-
37C, pH 7.8, mutant enzyme Q269stop
0.0101
tRNALeu
-
mutant lacking residues Q281 to D294, 45C
0.0114
tRNALeu
-
pH 8.2, 45C, mutant K139A
0.014
tRNALeu
Q15031
recombinant mitochondrial isozyme, pH 7.6, 37C
0.0174
tRNALeu
-
mutant enzyme L964K, at pH 8.2 and 30C
0.0273
tRNALeu
-
mutant enzyme L949K, at pH 8.2 and 30C
0.0003
tRNALeu from Aquifex aeolicus
-
aminoacylation reaction, pH 7.8, 60C
-
0.00038
tRNALeu from Aquifex aeolicus
-
aminoacylation reaction, pH 7.8, 37C
-
0.0014
tRNALeu from Aquifex aeolicus
-
recombinant enzyme complex, 55C
-
0.00076
tRNALeu from Escherichia coli
-
aminoacylation reaction, pH 7.8, 37C
-
0.0013
tRNALeu from Escherichia coli
-
recombinant enzyme complex, 55C
-
0.0015
tRNALeu from Escherichia coli
-
aminoacylation reaction, pH 7.8, 60C
-
0.00032
tRNALeu(GAG)
-
65C, recombinant His6-tagged enzyme
-
0.00045
tRNALeu(GAG)
-
65C, wild-type enzyme
-
0.0075
tRNALeu(GAG)
-
in 100 mM Tris-HCl (pH 7.8), 30 mM KCl, 12 mM MgCl2, 5 mM dithiothreitol, at 30C
-
0.0013
tRNALeu(UAA)
-
wild-type enzyme, aminoacylation
-
0.0013
tRNALeu(UAA)
-
pH 7.5, 37C, wild-type enzyme
-
0.0015
tRNALeu(UAA)
-
mutant enzyme T252V, aminoacylation
-
0.0017
tRNALeu(UAA)
-
mutant enzyme T252S, aminoacylation
-
0.0022
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T248V
-
0.0024
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247S/T248S
-
0.0026
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247V
-
0.0028
tRNALeu(UAA)
-
mutant enzyme T252A, aminoacylation
-
0.0036
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247A/T248A
-
0.0044
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247V/T248V
-
0.0076
tRNALeu(UAG)
-
in 100 mM Tris-HCl (pH 7.8), 30 mM KCl, 12 mM MgCl2, 5 mM dithiothreitol, at 30C
-
0.0179
tRNALeu(UUR)
-
-
-
0.0012
tRNALeuA35G
-
37C, pH 7.8
-
0.00052
tRNALeuA73
-
wild-type tRNALeu, pH 7.5, 37C
-
0.011
tRNALeuA73G
-
37C, pH 7.8
-
0.0002
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
0.0004
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
0.0017
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant L570F
-
0.0018
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutants K600L and K600R
-
0.002
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant L570R
-
0.0022
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant K600F
-
0.025
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant L570K
-
0.0003
tRNALeuGAG
-
pH 6.8, 65C, recombinant wild-type enzyme
-
0.0047
tRNALeuU73
-
tRNALeu isoacceptor, pH 7.5, 37C
-
0.000018
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant L570K
-
0.00016
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
0.0015
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant K600L
-
0.0017
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant L570F
-
0.004
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant K600R; human derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
0.006
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant K600F
-
0.0014
tRNASer mutant
-
pH 7.5, 37C
-
0.065
tubercidin 5'-triphosphate
-
-
0.01
Leu
-
ATP
additional information
additional information
-
-
-
additional information
additional information
-
-
-
additional information
additional information
-
kinetics, wild-type and mutant enzymes
-
additional information
additional information
-
substrate specificity with diverse tRNALeu isoacceptors and mutants
-
additional information
additional information
-
KM-values for hydrolytic editing of mischarged Ile-tRNALeu(GAG)
-
additional information
additional information
-
turnover numbers for tRNALeu(UUR) variants
-
additional information
additional information
-
kinetics of recombinant trunacted mutants, overview
-
additional information
additional information
-
kinetics of recombinant His-tagged wild-type and mutant enzymes
-
additional information
additional information
-
kinetics of chimeric mutants
-
additional information
additional information
-
kinetics of wild-type and mutant enzymes, overview
-
additional information
additional information
-
prolyl-tRNA synthetase, ProRS, and LeuRS interaction kinetics
-
additional information
additional information
-
kinetics of recombinant wild-type and mutant enzymes
-
additional information
additional information
-
kinetics of mischarging and post-transfer editing activities, overview
-
additional information
additional information
-
kcat/Km: 1 (ATP, wild-type), 1 (Leu, wild-type), 0.43 (ATP, mutant K587A), 0.42 (Leu, mutant K587A), 0.57 (ATP, mutant K588A), 0.37 (Leu, mutant K588A), 0.91 (ATP, mutant D603A), 0.94 (Leu, mutant D603A), 0.98 (ATP, mutant K606R), 0.99 (Leu, mutant K606R), 0.9 (ATP, mutant K606E), 0.99 (Leu, mutant K606E), 0.85 (ATP, mutant K606L), 0.98 (Leu, mutant K606L), 0.86 (ATP, mutant K606D), 0.83 (Leu, mutant K606D)
-
additional information
additional information
-
steady-state leucine activation and aminoacylation kinetics of GlLeuRS and its mutants, overview
-
additional information
additional information
-
equilibrium kinetics of wild-type and mutant enzymes, overview
-
additional information
additional information
-
kinetic constants of wild-type and D399A mutant of LeuRS in amino acid activation reaction with different amino acids, overview. Cytoplasmic LeuRS overexpressed in Escherichia coli exhibits the same kcat values as the one overexpressed in insect cells using in vitro transcribed tRNA
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.094
ATP
-
recombinant enzyme complex, 65C
0.22
ATP
-
recombinant mitochondrial isozyme mutant, 37C
0.27
ATP
-
37C, pH 7.6, leucylation, full-length enzyme
0.55
ATP
-
37C, pH 7.6, leucylation, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.74
ATP
-
37C, pH 7.6, ATP-diphosphate exchange, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.79
ATP
-
37C, pH 7.6, ATP-diphosphate exchange, full-length enzyme
0.8
ATP
Q15031
recombinant mitochondrial isozyme, pH 7.6, 37C
0.8
ATP
-
mutant E292K, pH 7.8, 37C
1.48
ATP
-
65C, wild-type enzyme
1.5
ATP
-
mutant E292S, pH 7.8, 37C
1.6
ATP
-
mutant enzyme, pH 7.8, 37C
1.8
ATP
-
aminoacylation reaction, pH 7.8, 60C
1.8
ATP
-
mutant E292F, pH 7.8, 37C
1.81
ATP
-
pH 6.8, 65C, recombinant wild-type enzyme
2
ATP
-
mutants E292Q and E292A, pH 7.8, 37C
2.2
ATP
-
mutant E292D, pH 7.8, 37C
2.2
ATP
-
65C, recombinant His6-tagged enzyme
3
ATP
-
pH 7.5, 65C, mutant K100A/Y105A
3.1
ATP
-
mutant lacking residues Q281 to D294, 45C; mutant lacking residues S295 to L304, 45C; wild-type, 45C
3.3
ATP
-
ATP-diphosphate exchange reaction, pH 7.8, 37C
3.6
ATP
-
wild-type enzyme, pH 7.8, 37C
3.6
ATP
-
recombinant enzyme
3.9
ATP
-
aminoacylation reaction, pH 7.8, 37C
4
ATP
-
pH 7.5, 65C, mutant F119A
4.2
ATP
-
native enzyme
4.2
ATP
-
pH 7.5, 65C, mutant V108A
4.3
ATP
-
pH 7.5, 65C, mutant D98A; pH 7.5, 65C, mutant E114A
4.6
ATP
-
pH 7.5, 65C, mutant K100A
4.7
ATP
-
pH 7.5, 65C, mutant D121A
4.8
ATP
-
37C, pH 7.8, mutant enzyme T25D
4.8
ATP
-
pH 7.5, 65C, mutant Y109A
4.9
ATP
-
wild-type enzyme, pH 7.8, 37C
4.9
ATP
-
37C, pH 7.8, mutant enzyme T252E
4.9
ATP
-
pH 7.5, 65C, mutant K100A/Y109A
5
ATP
-
wild-type enzyme, pH 7.8, 37C
5
ATP
-
37C, pH 7.8, native enzyme
5
ATP
-
pH 7.5, 65C, mutant T101A
5.1
ATP
-
pH 7.5, 65C, mutant N96A
5.4
ATP
-
pH 7.5, 65C, mutant R106A
5.5
ATP
-
pH 7.5, 65C, mutant R97A
5.8
ATP
-
pH 7.5, 65C, mutant E113A
6.5
ATP
-
pH 7.5, 65C, mutant Y105A
6.8
ATP
-
pH 7.5, 65C, wild-type enzyme
7.4
ATP
-
pH 7.5, 65C, mutant I104A; pH 7.5, 65C, mutant W103A
7.5
ATP
-
pH 8.2, 45C, mutant Q154A
7.7
ATP
-
pH 7.5, 65C, mutant T118A
8.1
ATP
-
pH 8.2, 45C, mutant K152A
13.8
ATP
-
pH 7.5, 65C, mutant I115A
14.5
ATP
-
ATP-diphosphate exchange reaction, pH 7.8, 60C
29.8
ATP
-
pH 8.2, 45C, mutant W155A
30.1
ATP
-
pH 8.2, 45C, mutant S153A
31
ATP
-
pH 8.2, 45C, mutant K170A
31.3
ATP
-
pH 8.2, 45C, mutant E165A
31.5
ATP
-
pH 8.2, 45C, mutant K141A
31.8
ATP
-
pH 8.2, 45C, mutant K144A
32.1
ATP
-
pH 8.2, 45C, mutant K148A
32.3
ATP
-
pH 8.2, 45C, mutant K166A
33.2
ATP
-
pH 8.2, 45C, mutant K139A
34.4
ATP
-
pH 8.2, 45C, mutant D173A
36.2
ATP
-
pH 8.2, 45C, mutant K142A
60.2
ATP
-
pH 8.2, 45C, mutant E167A
7
Ile-tRNALeu
-
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
7.4
Ile-tRNALeu
-
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
7.6
Ile-tRNALeu
-
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
8.8
Ile-tRNALeu
-
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
-
1.08
L-didehydroleucine
-
mutant T252Y
0.07
L-isoleucine
-
pH 7.5, 37C
0.1
L-isoleucine
-
pH 7.5, 37C
0.34
L-isoleucine
Q9P2J5
wild-type, pH 7.6, 30C
0.51
L-isoleucine
Q9P2J5
mutant D399A, pH 7.6, 30C
1.9
L-isoleucine
-
37C
6.9
L-isoleucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37C
10.2
L-isoleucine
P26637
-
18
L-isoleucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37C
0.19
L-isoleucyl-tRNALeu
-
mutant Y515A, pH 7.5, 37C
0.48
L-isoleucyl-tRNALeu
-
mutant Y520A, pH 7.5, 37C
0.93
L-isoleucyl-tRNALeu
-
mutant Y520H, pH 7.5, 37C
1.52
L-isoleucyl-tRNALeu
-
wild-type, pH 7.5, 37C
0.00961
L-leucine
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0.134
L-leucine
-
recombinant enzyme complex, 65C
0.18
L-leucine
-
recombinant mitochondrial isozyme mutant, 37C
0.23
L-leucine
-
recombinant mitochondrial isozyme mutant, 37C
0.3
L-leucine
-
37C, pH 7.6, leucylation, full-length enzyme
0.39
L-leucine
-
aminoacylation reaction, pH 7.8, 37C
0.56
L-leucine
-
37C, pH 7.6, leucylation, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.563
L-leucine
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0.8
L-leucine
-
mutant E292K, pH 7.8, 37C
0.8
L-leucine
-
mutant enzyme T252A, aminoacylation
0.81
L-leucine
-
37C, pH 7.6, ATP-diphosphate exchange, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.82
L-leucine
-
37C, pH 7.6, ATP-diphosphate exchange, full-length enzyme
1
L-leucine
-
mutant Y515E, pH 8.2, 45C
1.1
L-leucine
-
mutant Y520H, pH 8.2, 45C
1.28
L-leucine
-
-
1.4
L-leucine
-
aminoacylation reaction, pH 7.8, 60C
1.4
L-leucine
-
pH 6.8, 65C, recombinant wild-type enzyme
1.4
L-leucine
-
mutant Y520E, pH 8.2, 45C
1.5
L-leucine
-
mutant enzyme, pH 7.8, 37C
1.57
L-leucine
-
65C, wild-type enzyme
1.6
L-leucine
-
mutant E292S, pH 7.8, 37C
1.79
L-leucine
-
65C, recombinant His6-tagged enzyme
1.8
L-leucine
-
mutant E292F, pH 7.8, 37C
1.8
L-leucine
-
mutant Y515K, pH 8.2, 45C; mutant Y520A, pH 8.2, 45C
1.9
L-leucine
-
mutant E292A, pH 7.8, 37C
2
L-leucine
-
pH 7.5, 37C
2.1
L-leucine
Q9P2J5
wild-type, pH 7.6, 30C
2.2
L-leucine
-
mutant E292Q, pH 7.8, 37C
2.2
L-leucine
-
mutant T252Y
2.4
L-leucine
-
mutant E292D, pH 7.8, 37C
2.6
L-leucine
-
mutant Y515A, pH 8.2, 45C
2.7
L-leucine
Q15031
recombinant mitochondrial isozyme, pH 7.6, 37C
2.7
L-leucine
-
wild-type, pH 8.2, 45C
2.8
L-leucine
-
mutant lacking residues Q281 to D294, 45C; mutant lacking residues S295 to L304, 45C; wild-type, 45C
3
L-leucine
-
wild-type enzyme, pH 7.8, 37C
3
L-leucine
-
recombinant enzyme
3.1
L-leucine
-
pH 7.5, 65C, mutant E114A; pH 7.5, 65C, mutant K100A/Y105A
3.2
L-leucine
-
pH 7.5, 65C, mutant D98A
3.4
L-leucine
-
native enzyme
3.4
L-leucine
-
pH 7.5, 65C, mutant F119A
3.5
L-leucine
-
ATP-diphosphate exchange reaction, pH 7.8, 37C
3.8
L-leucine
-
pH 7.5, 65C, mutant T101A
3.9
L-leucine
-
pH 7.5, 65C, mutant K100A
4.1
L-leucine
-
pH 7.5, 65C, mutant V108A
4.3
L-leucine
-
pH 7.5, 65C, mutant Y105A
4.4
L-leucine
-
pH 7.5, 65C, mutant N96A
4.6
L-leucine
-
37C, pH 7.8, mutant enzyme T25D
4.6
L-leucine
-
pH 7.5, 65C, mutant K100A/Y109A
4.7
L-leucine
-
pH 7.5, 65C, mutant R106A; pH 7.5, 65C, mutant Y109A
4.9
L-leucine
-
37C, pH 7.8, mutant enzyme T252E
5.1
L-leucine
-
wild-type enzyme, pH 7.8, 37C
5.1
L-leucine
-
37C, pH 7.8, native enzyme
5.1
L-leucine
-
mutant enzyme T252S, aminoacylation
5.1
L-leucine
-
pH 7.5, 65C, mutant R97A; pH 7.5, 65C, mutant W103A
5.2
L-leucine
-
wild-type enzyme, pH 7.8, 37C
5.2
L-leucine
-
pH 7.5, 65C, mutant E113A
5.6
L-leucine
Q9P2J5
mutant D399A, pH 7.6, 30C
5.6
L-leucine
-
pH 7.5, 65C, mutant I104A
5.9
L-leucine
-
pH 8.2, 45C, mutant Q154A
6
L-leucine
-
pH 7.5, 65C, mutant D121A; pH 7.5, 65C, wild-type enzyme
6.1
L-leucine
-
wild-type enzyme, aminoacylation
6.2
L-leucine
-
mutant enzyme T252V, aminoacylation
6.2
L-leucine
-
pH 8.2, 45C, mutant K152A
8.1
L-leucine
-
pH 7.5, 65C, mutant T118A
10.7
L-leucine
-
pH 7.5, 65C, mutant I115A
11
L-leucine
-
pH 7.5, 37C
15.5
L-leucine
-
ATP-diphosphate exchange reaction, pH 7.8, 60C
25.8
L-leucine
-
pH 7.6, 37C, wild-type enzyme
26.2
L-leucine
-
pH 7.6, 37C, mutant D399A
26.3
L-leucine
-
37C
28.1
L-leucine
-
pH 8.2, 45C, mutant K170A
28.4
L-leucine
-
pH 8.2, 45C, mutant K148A
28.7
L-leucine
-
pH 8.2, 45C, mutant K166A
28.9
L-leucine
-
pH 8.2, 45C, mutant K144A
29.7
L-leucine
-
pH 8.2, 45C, mutant E165A
29.9
L-leucine
-
pH 8.2, 45C, mutant K139A
31.7
L-leucine
-
pH 8.2, 45C, mutant K142A
31.9
L-leucine
-
pH 8.2, 45C, mutant W155A
32.5
L-leucine
-
pH 8.2, 45C, mutant S153A
33.1
L-leucine
-
pH 8.2, 45C, mutant K141A
36.3
L-leucine
-
pH 8.2, 45C, mutant D173A
37.5
L-leucine
P26637
-
54.6
L-leucine
-
pH 8.2, 45C, mutant E167A
73.7
L-leucine
-
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
101
L-leucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37C
171
L-leucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37C
0.00119
L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.0418
L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
3.2
L-methionine
-
37C
7.6
L-methionine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37C
19
L-methionine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37C
12.6
L-norvaline
-
37C
0.06
L-oxonorvaline
-
mutant T252Y
2 - 3.7
Leu
-
aminoacylation
20 - 50
Leu
-
ATP-diphosphate exchange reaction
2.05
Leu-tRNALeu
-
-
3.15
Leu-tRNALeu
-
-
0.000418
Natrialba magadii tRNALeu(CAA)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.0464
Natrialba magadii tRNALeu(CAA)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.000668
Natrialba magadii tRNALeu(GAG)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
0.064
Natrialba magadii tRNALeu(GAG)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
-
13.5
norvaline
P26637
-
0.003
tRNALeu
-
37C, pH 7.8, mutant enzyme Q269stop
0.028
tRNALeu
-
37C, pH 7.6, tRNALeu from calf liver, leucylation, full-length enzyme
0.05
tRNALeu
-
mutant R449K
0.059
tRNALeu
-
37C, pH 7.6, tRNALeu from calf liver, leucylation, DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form)
0.072
tRNALeu
-
37C, pH 7.8, mutant enzyme N152A
0.12
tRNALeu
Q15031
recombinant mitochondrial isozyme, pH 7.6, 37C
0.12
tRNALeu
-
wild-type enzyme
0.177
tRNALeu
-
truncation mutant DELTA911-913, pH 7.5, 37C
0.2
tRNALeu
-
mutant W445Y
0.2
tRNALeu
-
pH 8.2, 45C, mutant K142A; pH 8.2, 45C, mutant K144A
0.31
tRNALeu
-
37C, pH 7.8, mutant enzyme R94A
0.32
tRNALeu
-
37C, pH 7.8, mutant enzyme V286stop; 37C, pH 7.8, wild-type enzyme
0.331
tRNALeu
-
full-length enzyme, pH 7.5, 37C
0.39
tRNALeu
-
37C, pH 7.8, mutant enzyme G237D; 37C, pH 7.8, mutant enzyme N163A
0.4
tRNALeu
-
37C, pH 7.8, mutant enzyme Q234H
0.4
tRNALeu
-
mutant V338A; mutant V338D; mutant V338E; mutant V338F; mutant V338L
0.42
tRNALeu
-
37C, pH 7.8, mutant enzyme K238A
0.47
tRNALeu
-
37C, pH 7.8, mutant enzyme M159A
0.5
tRNALeu
-
37C, pH 7.8
0.59
tRNALeu
-
37C, pH 7.8, mutant enzyme L283F
0.6
tRNALeu
-
mutant R451K
0.66
tRNALeu
-
37C, pH 7.8, mutant enzyme K160N
0.7
tRNALeu
-
pH 8.2, 45C, recombinant mutant DELTAESI/DELTAHsESI
0.8
tRNALeu
-
mutant E292K, pH 7.8, 37C
0.8
tRNALeu
-
pH 8.2, 45C, mutant D173A; pH 8.2, 45C, mutant K141A; pH 8.2, 45C, mutant Q154A
0.84
tRNALeu
-
37C, pH 7.8, mutant enzyme A156V
0.9
tRNALeu
-
pH 7.5, 37C, recombinant LS-domain deletion mutant
1
tRNALeu
-
pH 8.2, 45C, recombinant mutant T341A
1.1
tRNALeu
-
pH 8.2, 45C, mutant S153A
1.1
tRNALeu
-
pH 8.2, 45C, recombinant mutant D444A; pH 8.2, 45C, recombinant mutant T341R
1.2
tRNALeu
-
pH 8.2, 45C, mutant E167A
1.3
tRNALeu
-
mutant enzyme, pH 7.8, 37C
1.3
tRNALeu
-
pH 8.2, 45C, mutant K152A
1.3
tRNALeu
-
pH 8.2, 45C, recombinant mutant R338A
1.4
tRNALeu
-
pH 8.2, 45C, mutant K170A
1.5
tRNALeu
-
pH 8.2, 45C, mutant K139A
1.6
tRNALeu
-
mutant E292S, pH 7.8, 37C
1.8
tRNALeu
-
mutant E292A, pH 7.8, 37C
1.9
tRNALeu
-
mutant E292Q, pH 7.8, 37C
1.9
tRNALeu
-
pH 8.2, 45C, mutant K148A
2
tRNALeu
-
mutant E292F, pH 7.8, 37C
2.2
tRNALeu
-
mutant E292D, pH 7.8, 37C
2.4
tRNALeu
-
mutant enzyme V910P, at pH 8.2 and 30C
2.5
tRNALeu
-
pH 8.2, 45C, mutant W155A
2.7
tRNALeu
-
mutant lacking residues Q281 to D294, 45C; mutant lacking residues S295 to L304, 45C; wild-type, 45C
2.8
tRNALeu
-
pH 8.2, 45C, mutant E165A; pH 8.2, 45C, mutant K166A
2.9
tRNALeu
-
wild-type enzyme, pH 7.8, 37C
3.4
tRNALeu
-
recombinant enzyme
3.9
tRNALeu
-
native enzyme
4.7
tRNALeu
-
37C, pH 7.8, mutant enzyme T25D
5
tRNALeu
-
wild-type enzyme, pH 7.8, 37C
5
tRNALeu
-
wild-type enzyme
5
tRNALeu
-
mutant enzyme L949K, at pH 8.2 and 30C; mutant enzyme Q915K, at pH 8.2 and 30C
5.1
tRNALeu
-
wild-type enzyme, pH 7.8, 37C
5.1
tRNALeu
-
37C, pH 7.8, mutant enzyme T252E; 37C, pH 7.8, native enzyme
5.6
tRNALeu
-
mutant enzyme L949A, at pH 8.2 and 30C
7
tRNALeu
-
mutant enzyme V910A, at pH 8.2 and 30C
7.2
tRNALeu
-
mutant enzyme L964A, at pH 8.2 and 30C
7.8
tRNALeu
-
wild type enzyme, at pH 8.2 and 30C
9.2
tRNALeu
-
mutant enzyme V910W, at pH 8.2 and 30C
9.6
tRNALeu
-
pH 7.5, 37C, recombinant wild-type enzyme
10.6
tRNALeu
-
mutant enzyme L964K, at pH 8.2 and 30C; mutant enzyme R921K, at pH 8.2 and 30C
13.3
tRNALeu
-
mutant enzyme Q915A, at pH 8.2 and 30C
13.6
tRNALeu
-
mutant enzyme R921A, at pH 8.2 and 30C
0.006
tRNALeu from Aquifex aeolicus
-
recombinant enzyme complex, 55C
-
0.39
tRNALeu from Aquifex aeolicus
-
aminoacylation reaction, pH 7.8, 37C
-
1.5
tRNALeu from Aquifex aeolicus
-
aminoacylation reaction, pH 7.8, 60C
-
0.003
tRNALeu from Escherichia coli
-
recombinant enzyme complex, 55C
-
0.084
tRNALeu from Escherichia coli
-
aminoacylation reaction, pH 7.8, 37C
-
0.4
tRNALeu from Escherichia coli
-
aminoacylation reaction, pH 7.8, 60C
-
1.48
tRNALeu(GAG)
-
65C, wild-type enzyme
-
1.59
tRNALeu(GAG)
-
65C, recombinant His6-tagged enzyme
-
1.8
tRNALeu(GAG)
-
in 100 mM Tris-HCl (pH 7.8), 30 mM KCl, 12 mM MgCl2, 5 mM dithiothreitol, at 30C
-
0.02
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247V/T248V
-
0.1
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247A/T248A
-
0.8
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247V
-
0.9
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T248V
-
1.6
tRNALeu(UAA)
-
mutant enzyme T252A, aminoacylation
-
3.2
tRNALeu(UAA)
-
pH 7.5, 37C, wild-type enzyme
-
5.2
tRNALeu(UAA)
-
pH 7.5, 37C, mutant enzyme T247S/T248S
-
6.1
tRNALeu(UAA)
-
wild-type enzyme, aminoacylation
-
6.3
tRNALeu(UAA)
-
mutant enzyme T252V, aminoacylation
-
6.5
tRNALeu(UAA)
-
mutant enzyme T252S, aminoacylation
-
2
tRNALeu(UAG)
-
in 100 mM Tris-HCl (pH 7.8), 30 mM KCl, 12 mM MgCl2, 5 mM dithiothreitol, at 30C
-
0.31
tRNALeuA35G
-
37C, pH 7.8
-
0.11
tRNALeuA73G
-
37C, pH 7.8
-
0.35
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant K600L
-
0.39
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
3
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant K600F
-
3.2
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant K600R
-
9.8
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant L570K
-
14.5
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
21
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant L570R
-
24
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37C, recombinant mutant L570F
-
1.5
tRNALeuGAG
-
pH 6.8, 65C, recombinant wild-type enzyme
-
0.000018
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant L570K
-
0.00016
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
0.0017
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant L570F
-
0.003
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant K600L
-
0.09
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant wild-type enzyme
-
0.13
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant K600F
-
0.14
tRNALeuUUR
-
human derived substrate, pH 7.0, 37C, recombinant mutant K600R
-
23.8
Leu-tRNALeu
-
-
additional information
additional information
-
kinetics, mutant enzymes
-
additional information
additional information
-
turnover number for hydrolytic editing of mischarged Ile-tRNALeu(GAG)
-
additional information
additional information
-
turnover numbers for tRNALeu(UUR) variants
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
2.2
ATP
-
mutant lacking residues S295 to L304, 45C
4
3.6
ATP
-
mutant lacking residues Q281 to D294, 45C
4
4.5
ATP
-
wild-type, 45C
4
5.2
ATP
-
pH 7.5, 65C, mutant K100A/Y105A
4
6.4
ATP
-
pH 7.5, 65C, mutant I115A
4
6.6
ATP
-
pH 7.5, 65C, mutant T118A
4
6.9
ATP
-
pH 7.5, 65C, mutant E113A
4
7.3
ATP
-
pH 7.5, 65C, mutant F119A
4
8.1
ATP
-
pH 7.5, 65C, mutant Y109A
4
8.5
ATP
-
pH 7.5, 65C, mutant D121A
4
8.7
ATP
-
pH 7.5, 65C, mutant K100A
4
9.3
ATP
-
pH 7.5, 65C, mutant I104A
4
9.7
ATP
-
pH 8.2, 45C, mutant K152A
4
11.1
ATP
-
pH 8.2, 45C, mutant Q154A
4
11.5
ATP
-
pH 7.5, 65C, mutant E114A
4
11.8
ATP
-
pH 7.5, 65C, wild-type enzyme
4
11.9
ATP
-
pH 7.5, 65C, mutant D98A
4
12
ATP
-
pH 7.5, 65C, mutant V108A
4
12.1
ATP
-
pH 7.5, 65C, mutant Y105A
4
13.1
ATP
-
pH 7.5, 65C, mutant T101A
4
13.5
ATP
-
pH 7.5, 65C, mutant K100A/Y109A
4
13.8
ATP
-
pH 7.5, 65C, mutant N96A
4
15.7
ATP
-
pH 7.5, 65C, mutant W103A
4
17
ATP
-
pH 7.5, 65C, mutant R97A
4
18.2
ATP
-
pH 7.5, 65C, mutant R106A
4
26.8
ATP
-
pH 8.2, 45C, mutant E165A
4
28.3
ATP
-
pH 8.2, 45C, mutant E167A
4
33.5
ATP
-
pH 8.2, 45C, mutant D173A
4
38.7
ATP
-
pH 8.2, 45C, mutant K144A
4
40.8
ATP
-
pH 8.2, 45C, mutant K141A
4
40.9
ATP
-
pH 8.2, 45C, mutant K139A
4
42.3
ATP
-
pH 8.2, 45C, mutant S153A
4
45.4
ATP
-
pH 8.2, 45C, mutant K170A
4
45.6
ATP
-
pH 8.2, 45C, mutant W155A
4
46
ATP
-
pH 8.2, 45C, mutant K148A
4
47
ATP
-
pH 8.2, 45C, mutant K166A
4
49.7
ATP
-
pH 8.2, 45C, mutant K142A
4
3100
Ile-tRNALeu
-
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
0
3300
Ile-tRNALeu
-
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
0
3500
Ile-tRNALeu
-
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
0
3800
Ile-tRNALeu
-
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
0
0.16
L-isoleucine
Q9P2J5
wild-type, pH 7.6, 30C
311
0.17
L-isoleucine
Q9P2J5
mutant D399A, pH 7.6, 30C
311
2.7
L-isoleucine
-
37C
311
0.016
L-isoleucyl-tRNALeu
-
mutant Y515A, pH 7.5, 37C
6841
0.028
L-isoleucyl-tRNALeu
-
mutant Y520A, pH 7.5, 37C
6841
0.063
L-isoleucyl-tRNALeu
-
mutant Y520H, pH 7.5, 37C
6841
0.165
L-isoleucyl-tRNALeu
-
wild-type, pH 7.5, 37C
6841
0.0108
L-leucine
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
127
0.181
L-leucine
-
mutant Y520E, pH 8.2, 45C
127
0.185
L-leucine
-
mutant Y515E, pH 8.2, 45C
127
0.203
L-leucine
-
mutant Y520H, pH 8.2, 45C
127
0.225
L-leucine
-
mutant Y515K, pH 8.2, 45C
127
0.3
L-leucine
-
mutant Y520A, pH 8.2, 45C
127
0.442
L-leucine
-
wild-type, pH 8.2, 45C
127
0.481
L-leucine
-
mutant Y515A, pH 8.2, 45C
127
5.6
L-leucine
Q9P2J5
mutant D399A, pH 7.6, 30C
127
66.8
L-leucine
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
127
77.9
L-leucine
-
mutant lacking residues S295 to L304, 45C
127
94.3
L-leucine
Q9P2J5
wild-type, pH 7.6, 30C
127
186
L-leucine
-
mutant lacking residues Q281 to D294, 45C
127
286
L-leucine
-
pH 7.5, 65C, mutant F119A
127
311
L-leucine
-
wild-type, 45C
127
386
L-leucine
-
pH 7.5, 65C, mutant T101A
127
438
L-leucine
-
pH 7.5, 65C, mutant K100A
127
493
L-leucine
-
pH 7.5, 65C, mutant K100A/Y105A
127
498
L-leucine
-
pH 7.5, 65C, mutant E114A
127
521
L-leucine
-
pH 7.5, 65C, mutant Y109A
127
524
L-leucine
-
pH 7.6, 37C, mutant D399A
127
541
L-leucine
-
pH 7.5, 65C, mutant D98A
127
550
L-leucine
-
pH 7.5, 65C, mutant R106A; pH 7.5, 65C, mutant R97A
127
565
L-leucine
-
pH 7.6, 37C, wild-type enzyme
127
586
L-leucine
-
pH 7.5, 65C, mutant W103A
127
706
L-leucine
-
pH 7.5, 65C, mutant V108A
127
775
L-leucine
-
pH 7.5, 65C, mutant Y105A
127
806
L-leucine
-
pH 7.5, 65C, mutant K100A/Y109A
127
819
L-leucine
-
pH 7.5, 65C, mutant N96A
127
839
L-leucine
-
pH 7.5, 65C, mutant D121A
127
846
L-leucine
-
pH 7.5, 65C, mutant E113A
127
984
L-leucine
-
pH 7.5, 65C, mutant I104A
127
1013
L-leucine
-
pH 7.5, 65C, wild-type enzyme
127
1277
L-leucine
-
pH 7.5, 65C, mutant T118A
127
1686
L-leucine
-
pH 8.2, 45C, mutant Q154A
127
1889
L-leucine
-
pH 8.2, 45C, mutant K144A
127
1890
L-leucine
-
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37C
127
1892
L-leucine
-
pH 8.2, 45C, mutant E165A
127
1910
L-leucine
-
pH 7.5, 65C, mutant I115A
127
1952
L-leucine
-
pH 8.2, 45C, mutant K166A
127
1980
L-leucine
-
pH 8.2, 45C, mutant K139A
127
1993
L-leucine
-
pH 8.2, 45C, mutant K170A
127
2044
L-leucine
-
pH 8.2, 45C, mutant S153A
127
2186
L-leucine
-
pH 8.2, 45C, mutant K142A
127
2200
L-leucine
-
pH 8.2, 45C, mutant D173A
127
2290
L-leucine
-
pH 8.2, 45C, mutant K148A
127
2508
L-leucine
-
pH 8.2, 45C, mutant K141A
127
2583
L-leucine
-
pH 8.2, 45C, mutant K152A
127
2681
L-leucine
-
pH 8.2, 45C, mutant W155A
127
3370
L-leucine
-
pH 8.2, 45C, mutant E167A
127
9400
L-leucine
-
37C
127
0.0724
L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0
3.34
L-leucyl-Pyrococcus horikoshii tRNALeu(GAG)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0
3.3
L-methionine
-
37C
88
13
L-norvaline
-
37C
691
0.0318
Natrialba magadii tRNALeu(CAA)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0
4.21
Natrialba magadii tRNALeu(CAA)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0
0.199
Natrialba magadii tRNALeu(GAG)
-
isoform LeuRS2, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0
9.28
Natrialba magadii tRNALeu(GAG)
-
isoform LeuRS1, in 20 mM Tris-HCl, pH 9.0, 3.5 M KCl, 30 mM MgCl2, 1 mM dithiohtreitol, at 40C
0
38
tRNALeu
-
pH 8.2, 45C, mutant K144A
459
57
tRNALeu
-
pH 8.2, 45C, mutant K142A
459
73.2
tRNALeu
-
mutant lacking residues S295 to L304, 45C
459
75.3
tRNALeu
-
pH 8.2, 45C, recombinant mutant DELTAESI/DELTAHsESI
459
128.7
tRNALeu
-
mutant lacking residues Q281 to D294, 45C
459
132
tRNALeu
-
pH 8.2, 45C, mutant K139A
459
136
tRNALeu
-
pH 8.2, 45C, mutant D173A
459
157
tRNALeu
-
pH 8.2, 45C, mutant K141A
459
162.5
tRNALeu
-
pH 8.2, 45C, recombinant mutant R338A
459
178.6
tRNALeu
-
pH 8.2, 45C, recombinant mutant T341A
459
183.2
tRNALeu
-
mutant enzyme L949K, at pH 8.2 and 30C
459
186.4
tRNALeu
-
pH 8.2, 45C, recombinant mutant T341R
459
193
tRNALeu
-
pH 8.2, 45C, recombinant mutant D444A
459
210
tRNALeu
-
pH 8.2, 45C, mutant Q154A
459
245
tRNALeu
-
pH 8.2, 45C, mutant E167A
459
255
tRNALeu
-
pH 8.2, 45C, mutant K152A
459
260.9
tRNALeu
-
mutant enzyme V910P, at pH 8.2 and 30C
459
275
tRNALeu
-
pH 8.2, 45C, mutant K170A; pH 8.2, 45C, mutant S153A
459
337
tRNALeu
-
pH 8.2, 45C, mutant E165A
459
346
tRNALeu
-
pH 8.2, 45C, mutant K166A
459
352
tRNALeu
-
pH 8.2, 45C, mutant W155A
459
380
tRNALeu
-
pH 8.2, 45C, mutant K148A
459
442
tRNALeu
-
wild-type, 45C
459
609.2
tRNALeu
-
mutant enzyme L964K, at pH 8.2 and 30C
459
674.7
tRNALeu
-
mutant enzyme L949A, at pH 8.2 and 30C
459
2615
tRNALeu
-
mutant enzyme R921A, at pH 8.2 and 30C
459
3846
tRNALeu
-
mutant enzyme Q915K, at pH 8.2 and 30C
459
5889
tRNALeu
-
mutant enzyme R921K, at pH 8.2 and 30C
459
6000
tRNALeu
-
mutant enzyme L964A, at pH 8.2 and 30C
459
6364
tRNALeu
-
mutant enzyme V910A, at pH 8.2 and 30C
459
6571
tRNALeu
-
mutant enzyme V910W, at pH 8.2 and 30C
459
7000
tRNALeu
-
mutant enzyme Q915A, at pH 8.2 and 30C
459
7091
tRNALeu
-
wild type enzyme, at pH 8.2 and 30C
459
240
tRNALeu(GAG)
-
in 100 mM Tris-HCl (pH 7.8), 30 mM KCl, 12 mM MgCl2, 5 mM dithiothreitol, at 30C
0
260
tRNALeu(UAG)
-
in 100 mM Tris-HCl (pH 7.8), 30 mM KCl, 12 mM MgCl2, 5 mM dithiothreitol, at 30C
0
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.209
(2E)-3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)-1-phenylprop-2-en-1-one
-
pH 7.8, 37C
0.412
(2E)-3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-7-yl)-1-phenylprop-2-en-1-one
-
pH 7.8, 37C
1
(E)-[3-(1,3-dihydro-1-hydroxy-2,1-benzoxaborol-7-yl)]acrylic acid ethyl ester
-
above, pH 7.8, 37C
0.0323
1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl cyclohexylcarbamate
-
pH 7.8, 37C
0.0431
1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl phenylcarbamate
-
pH 7.8, 37C
0.0676
1-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-4-methylpentan-2-one
-
pH 7.8, 37C
0.0272
1-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]butan-2-one
-
pH 7.8, 37C
0.0308
1-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]pentan-2-one
-
pH 7.8, 37C
0.0221
2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.0309
2,1-benzoxaborole-1,6(3H)-diol
-
pH 7.8, 37C
0.0025
2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]pentan-3-one
-
pH 7.8, 37C
0.1
2-[3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]propyl]-1H-isoindole-1,3(2H)-dione
-
above, pH 7.8, 37C
0.005
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-3-methylbutan-2-one
-
pH 7.8, 37C
0.0045
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-4-methylpentan-2-one
-
pH 7.8, 37C
0.0038
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]butan-2-one
-
pH 7.8, 37C
0.0029
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]heptan-4-one
-
pH 7.8, 37C
0.0314
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]hexan-2-one
-
pH 7.8, 37C
0.0035
3-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]pentan-2-one
-
pH 7.8, 37C
0.0713
6-(2,2-dimethoxyethoxy)-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.1
6-(2-methoxyethoxy)-2,1-benzoxaborol-1(3H)-ol
-
above, pH 7.8, 37C
0.688
6-(3-hydroxypropyl)-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.0306
6-(benzyloxy)-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.1
6-(cyclohexylmethoxy)-2,1-benzoxaborol-1(3H)-ol
-
above, pH 7.8, 37C
0.0158
6-(propan-2-yloxy)-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.1
6-(pyridin-2-ylmethoxy)-2,1-benzoxaborol-1(3H)-ol
-
above, pH 7.8, 37C
0.063
6-(quinolin-2-yloxy)-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.0543
6-butoxy-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.013
6-ethoxy-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.0309
6-propoxy-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.0256
6-[(2-fluorobenzyl)oxy]-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.1
6-[(3-hydroxypentan-2-yl)oxy]-2,1-benzoxaborol-1(3H)-ol
-
above, pH 7.8, 37C
0.541
7-(3-hydroxypropyl)-2,1-benzoxaborol-1(3H)-ol
-
pH 7.8, 37C
0.578
ethyl (2E)-3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)prop-2-enoate
-
pH 7.8, 37C
0.0021
ethyl 2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]-2-methylpropanoate
-
pH 7.8, 37C
0.0016
ethyl 2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]butanoate
-
pH 7.8, 37C
0.0028
ethyl 2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]propanoate
-
pH 7.8, 37C
0.0167
ethyl 3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)propanoate
-
pH 7.8, 37C
1
ethyl 3-(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-7-yl)propanoate
-
above, pH 7.8, 37C
0.0417
ethyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy](phenyl)acetate
-
pH 7.8, 37C
0.0035
ethyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetate
-
pH 7.8, 37C
0.0036
methyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetate
-
pH 7.8, 37C
0.0777
N-ethyl-2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetamide
-
pH 7.8, 37C
0.1
N-tert-butyl-2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetamide
-
above, I114pH 7.8, 37C
0.0074
tert-butyl [(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetate
-
pH 7.8, 37C
0.1
tert-butyl [2-[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]ethyl]carbamate
-
above, pH 7.8, 37C
0.1
[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetaldehyde
-
above, pH 7.8, 37C
0.1
[(1-hydroxy-1,3-dihydro-2,1-benzoxaborol-6-yl)oxy]acetic acid
-
above, pH 7.8, 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.0068
-
-
0.011
-
recombinant mitochondrial isozyme mutant in cell lysate
0.024
-
mutant A293D, in crude enzyme extract of Escherichia coli strain TG1
0.047
-
purified recombinant wild-type enzyme, with substrate tRNALeu from Escherichia coli
0.05
-
recombinant mutant E292K, pH 7.8, 37C
0.12
-
recombinant mutant E292S, pH 7.8, 37C
0.128
-
purified recombinant mitochondrial isozyme mutant
0.13
-
mutant A293F, in crude enzyme extract of Escherichia coli strain TG1
0.14
-
mutants A293I, A293G, and A293Y, in crude enzyme extract of Escherichia coli strain TG1
0.14
-
recombinant mutants E292F and E292A, pH 7.8, 37C
0.16
-
recombinant mutant E292Q, pH 7.8, 37C
0.17
-
recombinant mutant E292D, pH 7.8, 37C
0.2
-
mutant A293R, in crude enzyme extract of Escherichia coli strain TG1
0.235
-
purified recombinant wild-type enzyme, with substrate tRNALeu from Aquifex aeolicus
0.27
-
mutant T252E, in crude enzyme extract of Escherichia coli strain TG1
0.28
-
wild-type enzyme, in crude enzyme extract of Escherichia coli strain TG1
0.29
-
mutant T252E/M328K, in crude enzyme extract of Escherichia coli strain TG1
0.3
-
mutant M328K, in crude enzyme extract of Escherichia coli strain TG1
0.35
-
recombinant wild-type enzyme, pH 7.8, 37C
0.79
-
recombinant CP1 domain mutant
1.233
-
-
1.3
-
purified recombinant wild-type enzyme, with substrate tRNALeu from Aquifex aeolicus
1.5
-
about, wild-type enzyme
1.5
-
purified recombinant wild-type enzyme, with substrate tRNALeu from Escherichia coli
1.54
-
purified recombinant enzyme
1.69
-
purified native enzyme
2.8
-
purified recombinant enzyme
additional information
-
activiy of several enzyme mutants and mixed mutants, containing subparts of the Escherichia coli enzyme
additional information
-
-
additional information
-
aminoacid activation activities of wild-type and mutant enzymes
additional information
-
-
additional information
-
activity and specificity of several truncation and deletion mutants, overview
additional information
-
-
additional information
-
hydrolytic post-transfer editing activity of wild-type and D345A mutant LeuRS CP1 domains, activities of truncation mutants, overview
additional information
-
-
additional information
-
mischarging and post-transfer editing activities, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 7
-
mitochondrial enzyme
6 - 9
-
little difference in activity within this range
6.8
-
assay at
7 - 7.5
-
assay at
7 - 8
-
cytoplasmic enzyme
7.4
-
aminoacylation assay at
7.5
-
in cacodylate(K+) buffer or HEPES buffer
7.5
-
assay at
7.5
-
assay at
7.5
Q72GM3
assay at
7.5
-
assay at
7.5
-
assay at
7.6
-
ATP-diphosphate exchange assay at
7.6
-
assay at, with HstRNALeu
7.6
-
assay at
7.8
-
ATP-diphosphate exchange assay at
7.8
-
assay at
7.8
-
assay at
7.9
-
assay at
8
-
aminoacylation assay at
8.2
-
assay at, with GltRNALeu
8.5
-
cytoplasmic enzyme
9
-
chloroplastic enzyme
9
-
in Tris-HCl or Bis-Tris-propane-HCl, the activity decreases obviously at pH 8.0 or below
9.5
-
in CHES-KOH, the activity decreases obviously at pH 8.0 or below
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
6 - 9
-
little difference in activity within this range
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
20
-
mitochondrial enzyme
22
-
assay at room temperature
25
-
assay at with the minihelix
30
-
assay at
30
P26637
assay at
35
-
cytoplasmic enzyme
37 - 65
-
broad temperature optimum
37
Q15031
assay at
37
-
assay at
37
-
assay at
37
-
assay at
37
-
assay at
37
-
assay at
37
-
assay at, with HstRNALeu
37
-
assay at
37
-
assay at
45
-
assay at, with GltRNALeu
65
-
misformation assay at
65
-
assay at
additional information
-
assay carried out at room temperature
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
5 - 40
-
5C: about 35% of maximal activity, 40C: about 45% of maximal activity, mitochondrial enzyme
20 - 40
-
20C: about 30% of maximal activity, 40C: about 85% of maximal activity, cytoplasmic enzyme
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
major part, enzyme participates in a multienzyme complex, not as large and stable as the one from nucleus
Manually annotated by BRENDA team
-
during restricted growth on low levels of leucine, localized in cytoplasmic membrane
-
Manually annotated by BRENDA team
-
predominantly
Manually annotated by BRENDA team
Q15031
mitochondrial isozyme
Manually annotated by BRENDA team
-
2.4% of total activity in the cell, enzyme participates in a large and stable multienzyme complex
Manually annotated by BRENDA team
-
during unrestricted growth, localized in this soluble protein
-
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Aquifex aeolicus (strain VF5)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Giardia intestinalis (strain ATCC 50803 / WB clone C6)
Mycoplasma mobile (strain ATCC 43663 / 163K / NCTC 11711)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Pyrococcus horikoshii (strain ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3)
Streptococcus pneumoniae (strain ATCC 700669 / Spain 23F-1)
Streptococcus pneumoniae (strain ATCC 700669 / Spain 23F-1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
33500
-
recombinant beta-subunit, gel filtration
652320
90000 - 100000
-
gel filtration, sucrose density gradient centrifugation
427
100000
-
gel filtration
432
100000
-
complexed recombinant leuS' and leuS gene product, i.e. alpha/beta-subunit complex, gel filtration
649605
100000
-
complexed recombinant leuS' and leuS gene product, gel filtration
651299
101000
Q15031
recombinant enzyme, gel filtration
650328
101900
-
calculation from nucleotide sequence
443
105000
-
gel filtration
274
107500
-
recombinant enzyme, gel filtration
652320
107600
-
electrospray mass spectrometry under native conditions
663318
116000
-
gel filtration
432
119000 - 120000
-
equilibrium sedimentation method
424
135000
-
gel filtration
435
135000
-
chloroplastic enzyme; gel filtration
437
135000
-
gel filtration
716384
145000
-
disc gel electrophoresis
426
150000
-
gel filtration
426
150000
-
HPLC gel filtration, component of the high-MW enzyme complex
439
200000
-
cytoplasmic enzyme, gel filtration
437
1200000
-
gel filtration, multienzyme complex
419
1200000
-
containing 9 amino acyl-tRNA synthetases specific for Glu, Pro, Ile, Leu, Gln, Lys, Arg and Asp; gel filtration, multienzyme complex
438
additional information
-
a series of molecular modeling studies including homology modeling and automated docking simulations are carried out. A 3D structure of Escherichia coli LeuRS is constructed via homology modling using the X-ray structure of Thermus thermophilus as a template because the LeuRS structure of Escherichia coli is not available from X-ray or NMR studies
663351
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
?
-
x * 129000 + x * ?, SDS-PAGE, the 129000 MW polypeptide is a component of the high-MW aminoacyl-tRNA synthetase complex (MW 1200000, gel filtration)
?
-
x * 55-60000, SDS-PAGE
?
-
prokaryotes and lower eukaryotes: the corresponding enzymes behave as free enzymes
?
-
the multienzyme complex is composed of 10 distinct polypeptides with MW ranging from 18000 to 165000
?
-
x * 129000, SDS-PAGE component of the high-MW multienzyme complex
?
-
nucleotide sequence
?
-
high molecular mass aminoacyl-tRNA synthetase complex with a coherent structure that can be visualized by electron microscopy
?
-
in mammalia and higher eukaryotes: multienzyme complex containing 9 amino acyl-tRNA synthetases specific for Glu, Pro, Ile, Leu, Gln, Lys, Arg and Asp
?
-
x * 135000, SDS-PAGE
?
-
x * 34000, N-terminal peptide after cleavage of peptide bond E292-A293, SDS-PAGE, x * 63000, C-terminal peptide after cleavage of peptide bond E292-A293, SDS-PAGE, x * 97300, recombinant wild-type enzyme, SDS-PAGE
?
-
x * 98500, recombinant enzyme, SDS-PAGE
?
-
the isolated CP1 domain of LeuRS is an active protein for editing mischarged tRNALeu(AAG). An insertion of 49 amino acid residues within the CP1 domain, the so-called 49-amino acid motif, is important for the optimal aminoacylation activity of LeuRS and is crucial for the editing capacity of LeuRS-CP1
?
-
x * 113000, calculated from sequence
?
-
x * 102200, isoform LeuRS1, calculated from amino acid sequence, x * 107600, isoform LeuRS2, calculated from amino acid sequence
?
-
x * 108000, calculated from amino acid sequence
?
-
x * 102200, isoform LeuRS1, calculated from amino acid sequence, x * 107600, isoform LeuRS2, calculated from amino acid sequence
-
?
-
x * 108000, calculated from amino acid sequence
-
dimer
-
2 * 63000, gel electrophoresis, after dissociation, the enzyme exhibits an equilibrium between an active dimeric form and an inactive monomeric form
dimer
-
heterodimer
dimer
-
1 * 74000, alpha-subunit, + 1 * 33500, beta-subunit, SDS-PAGE
dimer
-
enzyme exists as alpha beta dimer and as (alphabeta)2 tetramer
dimer
-
(alphabeta) heterodimer with a large extra domain, the leucine-specific domain, inserted into the catalytic domain, the subunit split site is exactly in the middle of the leucine-specific domain and may have a unique function, two peptides of eight and nine amino acid residues in the domain located in the alpha subunit are essential for the enzymes activity
monomer
-
1 * 120000, chloroplastic enzyme, SDS-PAGE of denatured enzyme
monomer
-
1 * 100000, SDS-PAGE
monomer
-
1 * 128000, SDS-PAGE
monomer
-
1 * 130000, cytoplasmic enzyme, SDS-PAGE of denatured enzyme
monomer
-
1 * 116000, SDS-PAGE
monomer
-
1 * 122000, urea-SDS-PAGE
monomer
-
1 * 116000, cytoplasmic enzyme, SDS-PAGE
monomer
-
1 * 100000, urea-SDS-PAGE
monomer
-
1 * 100000, chloroplastic enzyme, SDS-PAGE
monomer
Q15031
1 * 101000, recombinant enzyme, SDS-PAGE
tetramer
-
heterodimer in quartenary structure, 2 * 35000, leuS' gene product or beta-subunit, + 2 * 65000, leuS gene product or alpha-subunit, SDS-PAGE
tetramer
-
heterodimer in quartenary structure, 2 * 35000, leuS' gene product, + 2 * 65000, leuS gene product, SDS-PAGE
tetramer
-
enzyme exists as alpha beta dimer and as (alphabeta)2 tetramer
monomer
Cyberlindnera jadinii Torulopsis
-
1 * 128000, SDS-PAGE
-
additional information
-
class I enzyme with unique heterodimeric quartenary structure
additional information
-
determination od secondary structure elements in the enzyme and the isolated beta-subunit
additional information
-
determination of secondary structure elements in the wild-type and mutant enzyme
additional information
-
enzyme structure and tRNA binding site, the requirement of the C-terminal domain for misediting prevention is unique to LeuR, overview
additional information
-
primary and tertiary structure, overview, the RDW peptide, a highly conserved peptide within an RDW-containing motif, is important for enzyme interactions, overview
additional information
-
primary and tertiary structures of the LeuRS unique C-terminal domain, overview, the C-terminal extension of about 60 amino acids forms a discrete domain, which is unique among the LeuRSs and interacts with the corner of the L-shaped tRNALeu, overview
additional information
-
the amino acid editing site for LeuRS resides within the homologous CP1 domain: threonine-rich peptide and a second conserved GTG region that are separated by about 100 amino acids comprise parts of the hydrolytic editing site, comparison to IleRS, tertiary and primary structure analysis of the amino acid editing site, overview
additional information
-
homology modelling of Trypanosoma brucei LeuRS editing domain CP1 using Candida albicans LeuRS CP1 domain as the template, PDB ID 2WFG
additional information
-
three-dimensional LeuRS structure and sequence alignment of different eukaryal and archaeal LSD1s, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
proteolytic modification
-
processing of the mitochondrial isozyme by removing of the mitochondrial targeting presequence in the mitochondria of insect cells
lipoprotein
-
high molecular weight aminoacyl-tRNA synthetase complex contains lipid. Delipidation does not affect the size or activity of the complex, but a variety of functional and structural properties of individual synthetases in the complex are altered: sensitivity to salts plus detergents, temperature inactivation, hydrophobicity, sensitivity to protease digestion
additional information
-
the enzyme contains proteolytic cleavage sites at the peptide bonds between T252 and F253, E292 and A293, K327 and A328, and P368 and D369
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
structure at 2.2 A resolution of the editing domain in complex with compound AN3018, i.e.6-(ethylamino)-5-fluorobenzo[c][1,2]oxaborol-1(3H)-ol, using AMP as a surrogate for the 3' adenosine of tRNAleu. Comparison with the structure of the human enzyme
Q5A9A4
crystal and cocrystal structures for LeuRS, IleRS, and ValRS suggests that the CP1 domain rotates via its flexible beta-strand linkers relative to the main body along various steps in the enzymes reaction pathway. Computational analysis suggested that the end of the N-terminal beta-strand acted as a hinge. A molecular hinge might specifically direct movement of the CP1 domain relative to the main body
-
crystal structure of editing domain of Escherichia coli LeuRS in both apo form and complexes with methionine and isoleucine at 2.0 A, 2.4 A, and 3.2 A resolution, hanging drop vapour diffusion method
-
structure of the selenomethionine-labeled CP1 domain to 3.25 A, comparison with the structure of Candida albicans enzyme
-
hanging drop vapor diffusion method, using 0.1 M bis-Tris (pH 5.5), 0.6 M ammonium acetate, and 20% (w/v) PEG3350 at 4C
-
cocrystallizations with each of the tRNALeu isoacceptors are attempted. Cocrystals are obtained by the hanging-drop vapour-diffusion method, but only when the tRNALeu isoacceptor with the anticodon CAA is used. Electrophoretic analyses reveals that the crystals contain both leucyl-tRNA synthetase and tRNALeu, suggesting that they are LeuRS-tRNALeu complex crystals. A data set diffracting to 3.3 A resolution is collected from a single crystal at 100 K. The crystal belongs to the orthorhombic space group P2(1)2(1)2, with unit-cell parameters a = 118.18, b = 120.55, c = 231.13 A
-
hanging-drop vapour-diffusion method using ammonium sulfate as a precipitant. The crystals belong to the rhombohedral space group R3, with unit-cell parameters a = b = 186.20, c = 91.43 A, alpha = beta = 90, gamma = 120. The asymmetric unit contains one molecule of LeuRS, with a corresponding crystal volume per protein weight of 3.2 A3 Da(-1) and a solvent content of 60.7%. A data set diffracting to 2.2 A resolution is collected from a single crystal at -173C. Selenomethionine-substituted protein crystals are prepared in order to solve the structure by the SAD phasing method
-
hanging-drop vapour-diffusion method, resolution 2.1 A, C-terminally truncated LeuRS (amino acids 1-810), crystals belong to the Rhombohedral space group R3, with unit-cell parameters a = b = 186.23 A, c = 91.45 A, alpha = beta = 90, gamma = 120 for the native form and a = b = 185.8 A, c = 91.19 A, alpha = beta = 90, and gamma = 120 for the SeMet crystal
-
construction of a structural model of the completely solvated leucyl-tRNA synthetase complexed with valyl-tRNALeu
Q72GM3
crystal growth in presence of mercuric chloride, soaking of the crystals in solution containing 0.6 mM of the non-hydrolyzable substrate analogue norvaline-AMS for 1 month, or cocrystallization of enzyme and norvaline-AMS, X-ray diffraction structure determinationat 2.0-2.2 A resolution and analysis
Q72GM3
crystal structure of leucyl-tRNA synthetase complexed with tRNALeu in the post-transfer-editing conformation, crystals of the complex are grown at 20C by hanging drop vapour diffusion
-
crystallization of enzyme alone or in complex with leucine or leucyl-adenylate analogue, and crystallization of selenomethionine-enzyme, hanging-drop vapour-diffusion method with ammonium sulfate as precipitant, X-ray diffraction structure determination at 1.9-6 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
30
-
half-life: 4.5 min, in absence of 2-mercaptoethanol
426
40
-
2.5 min, 90% loss of activity of mitochondrial enzyme, 5-10 min, 90% loss of cytoplasmic enzyme
427
45
-
rapid inactivation
424
49.5
-
50% inactivation, heating of 0.5C per minute starting at 30C, mutant A293D
650024
54
-
50% inactivation, heating of 0.5C per minute starting at 30C, wild-type enzyme and mutants A293F, A293G, A293R
650024
55
-
50% inactivation, heating of 0.5C per minute starting at 30C, mutants A293I and A293Y
650024
65
-
stable up to under optimal conditions
652320
75
-
30 min, stable
652320
additional information
-
the mixed enzyme mutants are not thermostable in contrary to the wild-type enzyme
650221
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
extremely unstable in absence of 2-mercaptoethanol
-
unstable in the absence of sulfhydryl containing compound
-
20-30% loss of activity after 1 cycle of freezing and thawing
-
stability is lower with decreasing protein concentration
-
bovine serum albumin stabilizes
-
DTT stabilizes
-
DTT stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 10 mg/ml of enzyme, 1 month, partial cleavage of the enzyme into peptides of 63 and 34 kDA occurs, loss of 30% aminoacylation activity
-
4C, 50 mM potassium phosphate buffer, pH 7.0, 1 mM MgCl2, 0.002 mM L-leucine, 100 mM NaCl, 15% v/v 1,2-propanediol, stable for several weeks
-
-20C, 50% glycerol, stable for 2-3 years
-
15C, 0.1% Triton X-100, stable for several months
-
-20C, 50% glycerol, stable for several weeks
-
-20C, 50% glycerol, stable for at least 3 months
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
recombinant
-
recombinant enzyme and beta-subunit from Escherichia coli, to near homogeneity
-
recombinant enzymes from Escherichia coli, leuS' gene product to homogeneity
-
recombinant His-tagged alpha- and beta-subunits from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
single-step purification
-
Ni-NTA column chromatography
-
purified by affinity chromatography using His-select resin
-
recombinant enzyme
-
recombinant His-tagged enzyme from strain JM109(DE3), 4.6fold to homogeneity
-
recombinant His-tagged mutant and His-tagged CP1 domain expressed in Escherichia coli strain Jm109(DE3), to electrophoretic homogeneity
-
recombinant HIs-tagged wild-type and mutant enzymes
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from strain BL21(DE3) by nickel affiniry chromatography
-
recombinant His-tagged wild-type and mutant enzymes from strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from strain BL21(DE3) by nickel affinity chromatography to homogeneity
-
recombinant wild-type and mutant enzymes
-
recombinant wild-type and mutant enzymes from overproducing strain
-
recombinant wild-type enzyme from overexpression in Escherichia coli, purification of the proteolytically derived fragments from wild-type enzyme preparation
-
recombinant His-tagged GlLeuRS from Escherichia coli strain BL21-Codon Plus(DE3)-RIL by nickel affinity chromatography
-
recombinant protein
-
using Ni-NTA chromatography
-
recombinant enzyme
-
recombinant His-tagged LeuRS from Escherichia coli strain RosettaTM 2 (DE3) by nickel affinity and anion exchange chromatography, and ethanol precipitation, to 90% purity
-
recombinant N-terminally mutated His-tagged mitochondrial isozyme
-
using Ni-NTA chromatography
-
recombinant His6-tagged LeuRS from Escherichia coli strain BL21(DE3) by anion exchange chromatography, gel filtration, and ultrafiltration, co-purification of prolyl-tRNA synthetase, ProRS, and LeuRS, overview
-
Ni2+-NTA column chromatography
-
Ni-NTA column chromatography
-
Ni2+-NTA column chromatography, and gel filtration
-
wild-type enzyme, mutant enzyme found only in traces
-
recombinant C-terminally truncated LeuRS (amino acids 1-819)
-
recombinant His-tagged PhLeuRS from Escherichia coli strain BL21-Codon Plus(DE3)-RIL by nickel affinity chromatography
-
purified by affinity chromatography using His-select resin
-
recombinant from Escherichia coli
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography to homogeneity
-
using Ni-NTA chromatography
P26637
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
2 genes leuS and leuS', overexpression in Escherichia coli BL21(DE3)
-
DNA fragment encoding the fusion protein of Escherichia coli CP1 domain with Aquifex aeolicus beta-subunit and CP1 Escherichia coli beta-like polypeptide are inserted into pET30a by a two step procedure
-
Escherichia coli BL21-Codon Plus (DE3) cells are co-transformed with plasmid pSBETH6-lrsa and pET-15b-lrsb to produce alphabeta-LeuRS with an N-terminal His6 tag
-
expression in Escherichia coli
-
expression of alpha subunit and beta subunit in Escherichia coli
-
expression of the enzyme and the beta-subunit alone in Escherichia coli, the alpha-subunit cannot be expressed stably
-
individual expression of His-tagged alpha- and beta-subunits in Escherichia coli strain BL21(DE3)
-
overexpression in Escherichia coli
-
overexpression of the alpha and beta subunits and the C- and N-terminal parts of the Escherichia coli enzyme in Escherichia coli as monomeric and dimeric mutants, also mixed between the species
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expressed in Escherichia coli BL21(DE3) cells
-
expression and assembly of the 63 and 34 kDa peptides in Escherichia coli strain KL231 cannot complement the deficient temperature-sensitive strain, while expression of peptide fragments derived by cleavage of other peptide bonds leads to assembly of a functional enzyme
-
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
-
expression of His-tagged wild-type and mutant enzymes in strain BL21(DE3), subcloning in strain DH5alpha
-
expression of His-tagged wild-type and mutants in Escherichia coli strain SG13009
-
expression of the His-tagged CP1 domain mutant and isolated CP1 domain in Escherichia coli strain JM109(DE3)
-
expression of wild-type and mutants in Escherichia coli strain TG1
-
gene leuS, expression in a BL21 strain
-
gene leuS, expression of His-tagged wild-type and mutant enzymes in strain BL21(DE3)
-
gene leuS, overexpression in strain JM109(DE3) as His-tagged protein
-
overexpression of the N-terminal and the C-terminal parts of the enzyme and of the alpha- and beta-subunits of the Aquifex aeolicus enzyme in an Escherichia coli strain as monomeric and dimeric mutants, also mixed between the species
-
overexpression of wild-type and mutants
-
subcloning in strain DH5alpha, expression of His-tagged wild-type and mutant enzymes in strain BL21(DE3)
-
the gene fragment encoding the editing domain (residues 228-413) is subcloned into the pET3E-His expression plasmid. The plasmid is transformed into the Escherichia coli BL-21(DE3) strain
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expression in Escherichia coli
-
expression in Escherichia coli strain KL231 cells
-
overexpression of His-tagged GlLeuRS in Escherichia coli strain BL21-Codon Plus(DE3)-RIL. Complementation assay of the thermosensitive Escherichia coli KL231 strain
-
in vitro transcription
-
expressed in Escherichia coli as a His-tagged fusion protein
-
expression of hcLeuRS and DELTAChcLeuRS (a C-terminal 89-amino acid truncated enzyme form) in a baculovirus system
-
expression of His-tagged cytoplasmic LeuRS in Escherichia coli strain RosettaTM 2 (DE3), expression in Spodotera frugiperda cells via baculovirus transfection system
-
gene LARS1
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mitochondrial isozyme, DNA sequence determination and analysis, expression of the mature enzyme as His-tagged enzyme in Escherichia coli BL21(DE3)
Q15031
the mitochondrial isozyme is cloned and expressed in Escherichia coli with the same N-terminus as that processed in the mitochondria of insect cells as His-tagged protein
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expression of prolyl-tRNA synthetase, ProRS, and LeuRS in a Saccharomyces cerevisiae two-hybrid system, expression of His6-tagged LeuRS in Escherichia coli strain BL21(DE3)
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21(DE3) cells
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expressed in Escherichia coli BL21(DE3) cells
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expression in Escherichia coli
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overexpressed in a C-terminally truncated form in Escherichia coli
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overexpression of His-tagged PhLeuRS in Escherichia coli strain BL21-Codon Plus(DE3)-RIL. Complementation assay of the thermosensitive Escherichia coli KL231 strain
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expressed in Escherichia coli as a His-tagged fusion protein
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expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
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expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3), complementation abilities of wild-type and mutant enzymes of yeast null strain HM402 and Escherichia coli strain KL321, overview
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gene CDC60, expression of wild-type and mutants in an Escherichia coli BL21 strain
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LeuRS expression in Escherichia coli strain BL21(DE3)-RIPL
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
A156V
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mutation in beta-subunit, the ratio of turnover-number to Km-value is identical to the wild-type ratio
D373A
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mutant defective in post-transfer editing function
G237D
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 1.7fold higher than the wild-type ratio
K238A
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 2.4fold higher than the wild-type ratio
K599A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K599A/K600A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K599A/K600A/K605A/K606A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K600A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K605A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K605A/K606A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
K606A
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site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity
L283F
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 2fold higher than the wild-type ratio
M159A
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 80% of the wild-type value
N152A
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 16% of the wild-type ratio
N163A
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 2.4fold higher than the wild-type ratio
Q234H
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mutation in beta-subunit, the ratio of turnover-number to Km-value is identical to the wild-type ratio
Q260stop
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 0.02% of the wild-type ratio
R94A
-
mutation in beta-subunit, the ratio of turnover-number to Km-value is 40% of the wild-type ratio
T273R
-
no change in aminoacylation activity, but the deacylation of Ile-tRNALeu is strongly impaired. Mutant still exhibits 70% of wild-type AMP formation
ts025C1
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temperature sensitive mutant ts025C1
tsH1
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temperature sensitive mutant tsH1
A293D
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81% decreased activity, highly decreased editing function, very strong binding of ATP, high decrease in Km for the substrates, more sensitive too inhibition by ATP, increased heat lability compared to the wild-type
A293D
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site-directed mutagenesis, the mutant activity is similar to the wild-type enzyme
A293E
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site-directed mutagenesis, the mutant activity is similar to the wild-type enzyme
A293F
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54% decreased activity compared to the wild-type, more sensitive too inhibition by ATP
A293G
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50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
A293I
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51% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
A293K
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site-directed mutagenesis, the post-transfer editing activity of the isolated CP1-domain is enhanced compared to the wild-type enzyme's domain
A293R
-
30% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates
A293R
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site-directed mutagenesis, the post-transfer editing activity of the isolated CP1-domain is enhanced compared to the wild-type enzyme's domain
A293Y
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50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
D251W
-
site-directed mutagenesis, editing site mutant, the substrate specificity and charging fidelity is retained
D345A
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mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine
D345A
-
site-directed mutagenesis, the mutation in the isolated CP1-domain abolishes hydrolytic post-transfer editing activity
DELTA788-798
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partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. This mutant shows almost no aminoacylation activity. Mutant shows reduced deacylation activity against mischarged Ile-tRNALeu
DELTA790-798
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partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant shows reduced deacylation activity against mischarged Ile-tRNALeu
DELTA792-798
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partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
DELTA793
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single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA794
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single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA794-798
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partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
DELTA795
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single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA795-796
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two-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA795-798
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partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
DELTA796
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single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA796-798
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partial deletion of the C-terminal domain peptide linker shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu, two-site deletion at the more flexible end of the peptide linker: mutant exhibits lower aminoacylation activity compared to wild-type
DELTA797
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single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA797-798
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two-site deletion at the more flexible end of the peptide linker: mutant exhibits lower aminoacylation activity compared to wild-type
E184A
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the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
E184R
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the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme. The substitution specifically inhibits tRNA-dependent pre-transfer editing
E184R/T252R
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the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
E292A
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unaltered specific activity in amino acid activation reaction, 61% reduced aminoacylation activity compared to the wild-type
E292D
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unaltered specific activity in amino acid activation reaction, 53% reduced aminoacylation activity compared to the wild-type
E292F
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unaltered specific activity in amino acid activation reaction, 60% reduced aminoacylation activity compared to the wild-type
E292K
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unaltered specific activity in amino acid activation reaction, 85% reduced aminoacylation activity compared to the wild-type
E292Q
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unaltered specific activity in amino acid activation reaction, 54% reduced aminoacylation activity compared to the wild-type
E292S
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unaltered specific activity in amino acid activation reaction, 66% reduced aminoacylation activity compared to the wild-type
E797GGG
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mutant shows no altered aminoacylation activity compared to wild-type
E797PPP
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mutant shows no altered aminoacylation activity compared to wild-type
G225P
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abolishes tRNA leucylation due to a defect in leucine activation, decrease in deacylation of Ile-tRNALeu
G229P
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increased aminoacylation activity compared to the wild-type, mutant deacylates Ile-tRNALeu similar to wild-type
G229P/T252A
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double mutant rescues leucylation activity to levels comparable to wild-type and retains deacylation activity of LeutRNALeu that is characteristic of the T252A mutation
G407P
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aminoacylates tRNALeu and decylates Ile-tRNALeu as well as wild-type
G409P
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increased aminoacylation activity compared to the wild-type, mutant deacylates Ile-tRNALeu similar to wild-type
G409P/T252A
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double mutant fails to rescue the T252A mutation in LeuRS
K809A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K846A
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
K846A/K853A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K846E
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site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
K846E/K853E
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K853A
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site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-typ enzyme
K853E
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
L570F
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the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, it shows a catalytic turnover for isoleucine decreased by a factor of 2, L570F has an 11fold higher Km for leucine compared to the wild-type enzyme, the activity is reduceDdcompared to the wild-type enzyme
L570K
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the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, it shows a catalytic turnover for isoleucine decreased by a factor of 2, L570F has an 11fold higher Km for leucine compared to the wild-type enzyme, the activity is reduced compared to the wild-type enzyme
L570R
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the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, it shows a catalytic turnover for isoleucine decreased by a factor of 2, L570R has a 4fold stronger binding affinity for leucine compared to the wild-type enzyme, the activity is reduce compared to the wild-type enzyme
L854A
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
L855A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
M328K
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7% increased activity compared to the wild-type
M336A
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site-directed mutagenesis, editing site mutant, the mutant shows a small increase in leucine editing activity
N807A
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site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
N807A/N856A
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site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
N856A
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site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
Q805A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
Q805A/N807A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
Q805A/N807A/N856A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
R185E
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the mutation significantly enhances tRNA-dependent pre-transfer editing activity
R249F
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site-directed mutagenesis, editing site mutant, editing activity of Leu-tRNALeu is decreased
R249F/T252A
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site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity
R249T
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site-directed mutagenesis, editing site mutant, the mutant shows increased activity with tRNALeu, but even higher activity with tRNAIle compared to the wild-type enzyme
R249T/D251W
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site-directed mutagenesis, editing site mutant, the mutant shows decreased hydrolysis of mischarged Ile-tRNALeu compared to the wild type enzyme
R286E
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the mutation significantly enhances tRNA-dependent pre-transfer editing activity
R811A
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site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
T247A/T248A
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533fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247S/T248S
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77fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V
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8fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V
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site-directed mutagenesis, hydrolysis of Ile-tRNALeu is completely abolished
T247V/T248V
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fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V/T248V
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site-directed mutagenesis, the double mutation abolishes post-transfer editing activity
T248V
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6fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T248V
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site-directed mutagenesis, hydrolysis of Ile-tRNALeu is completely abolished
T252A
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decreased activity with L-leucine, mutant shows altered editing specificity, it edits correctly formed leucyl-tRNALeu
T252A
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the ratio of turnover-number to KM-value for L-leucine in aminoacylation is 5% of the wild-type ratio, the ratio of turnover-number to KM-value for tRNALeu in aminoacylation is 10% of the wild-type ratio. The ratio of turnover-number to Km value for Leu-tRNALeu(UAA) is 25fold higher than the wild-type value, the ratio of turnover-number to Km value for Ile-tRNALeu(UAA) is 1.25fold higher than the wild-type value
T252A
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site-directed mutagenesis, the mutation in the full-length LeuRS uncouples specificity and hydrolyzes correctly charged LeutRNALeu
T252A
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site-directed mutagenesis, the mutation uncouples specificity and shows a 24-fold increase in hydrolytic activity compared to the wild-type enzyme, introduction of the large aromatic residue at Arg249 or Val338 rescued leucylation activity of the T252A mutation
T252D
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mutation results in isoleucylation of tRNALeu, editing activity is impaired, ATP hydrolysis in presence of norvaline is 27% of the wild-type value, ATP hydrolysis in presence of leucine is 98% of the wild-type value
T252E
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activity is similar to the wild-type
T252E
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mutation results in isoleucylation of tRNALeu, editing activity is impaired, ATP hydrolysis in presence of norvaline is 18% of the wild-type value, ATP hydrolysis in presence of leucine is 86% of the wild-type value
T252E/M328K
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activity is similar to the wild-type
T252F
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impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine
T252G
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the mutant enzyme, like the native enzyme, does not mischarge tRNALeu with isoleucine, ATP hydrolysis in presence of norvaline is 2.1fold higher than wild-type value, ATP hydrolysis in presence of leucine is 60% of the wild-type value
T252L
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impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine
T252R
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the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
T252S
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the ratio of turnover-number to KM-value for L-leucine in aminoacylation is 60% of the wild-type ratio, the ratio of turnover-number to KM-value for tRNALeu in aminoacylation is 80% of the wild-type ratio. The ratio of turnover-number to Km value for Leu-tRNALeu(UAA) is 7.5fold higher than the wild-type value, the ratio of turnover-number to Km value for Ile-tRNALeu(UAA) is 1.6fold lower than the wild-type value
T252V
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the ratio of turnover-number to KM-value for L-leucine in aminoacylation is identical to wild-type ratio, the ratio of turnover-number to KM-value for tRNALeu in aminoacylation is 90% of the wild-type ratio. The ratio of turnover-number to Km value for Leu-tRNALeu(UAA) is 1.75fold higher than the wild-type value, the ratio of turnover-number to Km value for Ile-tRNALeu(UAA) is 13.3fold higher than the wild-type value
T252Y
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impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine, effective aminoacylation of tRNALeu with allylglycine, homopropargylglycine, 2-butynylalanine, norvaline, and norisoleucine
T252Y
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site-directed mutagenesis, the mutation occupies the amino acid binding pocket and blocks the binding of substrate to abolish editing activity
T252Y
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mutant is unable to proofread amino acids with unbranched side chains, and enables insertion of a variety of noncanonical amino acids into recombinant proteins in place of leucine
T272R
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no change in aminoacylation activity, but the deacylation of Ile-tRNALeu is strongly impaired. Mutant still exhibits 45% of wild-type AMP formation
V338A
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site-directed mutagenesis, editing site mutant, it shows increased post-transfer editing activity of Leu-tRNALeu compared to the wild-type enzyme
V338D
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site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
V338E
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site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
V338F
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site-directed mutagenesis, editing site mutant, single introduction of the bulky phenylalanine residue nearly abolished post-transfer editing activity and facilitated mischarging of both isoleucine and valine to tRNALeu, 3000fold reduced activity
V338F/T252A
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site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity
V338L
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site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
D173A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
D444A
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site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D444A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
D444E
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site-directed mutagenesis
D588A
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kcat/Km: 0.57 (ATP), 0.37 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type
D603A
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kcat/Km: 0.91 (ATP), 0.94 (Leu), similar activity compared to wild-type
DELTA581-617
-
deletion of the CP2 domain, an insertion domain called connective peptide 2, of Giardia lamblia LeuRS shows that the CP2 domain is indispensable for amino acid activation, post-transfer editing and contributes to LeuRS-tRNALeu binding affinity. CP2 domain of Pyrococcus horikoshii LeuRS but not that of Escherichia coli LeuRS can partially restore amino acid activation and post-transfer editing functions suggesting that the functions of the CP2 domain are dependent on its location in the primary sequence of LeuRS
E165A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
E167A
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site-directed mutagenesis, the mutant shows defects in leucine activation, mutant kinetics compared to the wild-type enzyme, overview
E298A
-
activity similar to wild-type
F171A
-
site-directed mutagenesis, inactive mutant
K139A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K141A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K142A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K144A
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site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K148A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K152A
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site-directed mutagenesis, the mutant shows defects in leucine activation, mutant kinetics compared to the wild-type enzyme, overview
K166A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K170A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
K299A
-
activity similar to wild-type
K303A
-
activity similar to wild-type
K587A
-
kcat/Km: 0.43 (ATP), 0.42 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type
K606A
-
mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities
K606D
-
kcat/Km: 0.86 (ATP), 0.83 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type
K606E
-
kcat/Km: 0.90 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type
K606L
-
kcat/Km: 0.85 (ATP), 0.98 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type
N301A
-
activity similar to wild-type
Q154A
-
site-directed mutagenesis, the mutant shows defects in leucine activation, mutant kinetics compared to the wild-type enzyme, overview
R338A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
S153A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
S295A
-
activity similar to wild-type
T341A
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
T341R
-
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
W155A
-
site-directed mutagenesis, mutant kinetics compared to the wild-type enzyme, overview
W586A
-
mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities
Y515A
-
109% of wild-type activity
Y515E
-
42% of wild-type activity
Y515K
-
51% of wild-type activity
Y520A
-
68% of wild-type activity
Y520E
-
41% of wild-type activity
Y520H
-
46% of wild-type activity
Y581A
-
mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities
Y581E
-
mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity
Y581F
-
mutant shows some but significantly reduced leucine activation activity its post-transfer editing activity is similar to wild-type
Y581K
-
mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity
Y581S
-
mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity
A3243G
-
respiratory chain defects in A3243G mutant cells is suppressed by overexpressing human mitochondrial leucyl-tRNA synthetase. The rates of oxygen consumption in suppressed cells are directly proportional to the levels of leucyl-tRNA synthetase. 15fold higher levels of leucyl-tRNA synthetase results in wild-type respiratory chain function. The suppressed cells have increased steady-state levels of tRNA(Leu(UUR)) and up to 3fold higher steady-state levels of mitochondrial translation products, but do not have rates of protein synthesis above those in parental mutant cells
D399A
Q9P2J5
40fold increase in Km for leucine activation. Mutation eliminates Ile-tRNALeu deacylation activity
D399A
-
site-directed mutagenesis, tRNA selectivity compared to the wild-type enzyme
D399K
-
mutant is resitant to inhibitor 5-fluoro-2,1-benzoxaborol-1(3H)-ol but more sensitive to norvaline inhibition
K600F
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, the mutant demonstrates a 9fold decrease in its ability to distinguish between leucine and isoleucine effectively, the activity is reduced compared to the wild-type enzyme
K600L
-
the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, the mutant demonstrates an 11fold increase in its ability to distinguish between leucine and isoleucine effectively, the activity is reduced compared to the wild-type enzyme
T298A
Q9P2J5
activity similar to wild-type, mutation maintains Ile-tRNALeu deacylation activity
T298Y
Q9P2J5
mutation uncouples specificity in the editing active site and mutant hydrolyzes Leu-tRNALeu
L949A
-
the mutant shows 9.5% activity compared to the wild type enzyme
L949K
-
the mutant shows 2.6% activity compared to the wild type enzyme
L964A
-
the mutant shows 85% activity compared to the wild type enzyme
L964K
-
the mutant shows 8.6% activity compared to the wild type enzyme
Q915A
-
the mutant shows 99% activity compared to the wild type enzyme
Q915K
-
the mutant shows 54% activity compared to the wild type enzyme
R921A
-
the mutant shows 37% activity compared to the wild type enzyme
R921K
-
the mutant shows 83% activity compared to the wild type enzyme
V910A
-
the mutant shows 90% activity compared to the wild type enzyme
V910P
-
the mutant shows 3.7% activity compared to the wild type enzyme
V910W
-
the mutant shows 93% activity compared to the wild type enzyme
Q915A
-
the mutant shows 99% activity compared to the wild type enzyme
-
Q915K
-
the mutant shows 54% activity compared to the wild type enzyme
-
V910A
-
the mutant shows 90% activity compared to the wild type enzyme
-
V910P
-
the mutant shows 3.7% activity compared to the wild type enzyme
-
V910W
-
the mutant shows 93% activity compared to the wild type enzyme
-
D121A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
D332A
-
site-directed mutagenesis
D332A
-
unlike the wild-type enzyme the mutant enzyme synthesizes the incorrect product Ile-tRNALeu, unlike the wild-type enzyme, the mutant enzyme cannot deacylate Ile-tRNALeu
D98A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
E113A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
E114A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
F119A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
I104A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
I115A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K100A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
K100A/Y105A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
N96A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
R106A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
R97A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
T101A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
T118A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
V108A
-
site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
W103A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
Y105A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
Y109A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
D357A
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site-directed mutagenesis, the mutant shows reduced activity and abolished editing activity and misaminoacylated isoleucine to tRNALeu compared to the wild-type enzyme
D418R
P26637
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
D419A
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mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine
D419A
P26637
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Mutant D419A shows mischarging capacity with Ile. Growth rate is severly inhibited in medium containing a large excess of norvaline and reduced leucine
DELTA270-530
P26637
deletion of the CP1 domain shows that the mutant is not able to rescue LeuRS knock-out strain
DELTA314-319
P26637
deletion of the T-rich region shows that the mutant is able to rescue LeuRS knock-out strain with a grwoth rate similar to wild-type
DELTA819-828
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deletion of the C-terminal domain peptide linker stimulates aminoacylation and editing activity shows that as the length of the peptide linker decreases, aminoacylation activity decreases. Mutant retains significant deacylation activity against mischarged Ile-tRNALeu
R265A
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site-directed mutagenesis, the mutant shows reduced activity and abolished post-transfer editing activity compared to the wild-type enzyme
R449A
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site-directed mutagenesis, nearly inactive mutant
R449E
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
R449K
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320, 30fold reduced activity compared to the wild-type enzyme
R451A
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site-directed mutagenesis, nearly inactive mutant
R451E
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
R451K
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site-directed mutagenesis, mutation within the RDW peptide, complementation of the null mutant strain QBY320, 11fold reduced activity compared to the wild-type enzyme
S416D
P26637
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
T263V/T264V
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site-directed mutagenesis, the mutant shows reduced activity and decreased post-transfer editing activity compared to the wild-type enzyme
T31E
P26637
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Mutant T319A shows mischarging capacity with Ile. Growth rate is severly inhibited in medium containing a large excess of norvaline and reduced leucine
T347A
P26637
mutant is inhibited by AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
T410A
P26637
mutant is inhibited by AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
W445A
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site-directed mutagenesis, nearly inactive mutant
W445F
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445H
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445K
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site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445Y
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site-directed mutagenesis, mutation within the RDW peptide, weak complementation of the null mutant strain QBY320, 30fold reduced activity compared to the wild-type enzyme
M160N
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 2.4fold higher than the wild-type ratio
additional information
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deletion of the last 36 residues of the alpha-subunit is deleterious for tRNA charging, resulting in an inactive mutant, the monomeric mutants containing the alpha-subunit have activities comparable to the wild-type enzyme, while the heterodimeric enzymes show very low activity
additional information
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construction of diverse truncation mutants, ATP-diphosphate-exchange and aminoacylation activities, and ability to charge minihelixLeu of alpha and beta subunit mutants, overview
additional information
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the freestanding LeuRS editing domain can edit this precursor in contrast to IleRS and ValRS editing domains, overview, design and preparation of minihelixLIV, overview
V286stop
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mutation in beta-subunit, the ratio of turnover-number to Km-value is 0.6% of the wild-type ratio
additional information
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a mutation in the gene, leuS1, increases the transcription and expression of the ilv-leu operon, permitting monitoring of leuS alleles
additional information
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using a Chinese hamster ovary cell line containing a temperature-sensitive mutation in leucyl-tRNA synthetase which is active at 34 C but defective at 39.5 C, it is shown that shifting the cells to the latter temperature mimics the effects of amino acid starvation on protein synthesis. Leucine deprivation markedly inhibits mTORC1 signaling in these cells, but shifting the cells to the nonpermissive temperature for the synthetase does not. These data indicate that uncharged tRNALeu does not switch off mTORC1 signaling and suggest that mTORC1 is controlled by a distinct pathway that senses the availability of amino acids
M336F/T252A
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site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity, M336F/T252A double LeuRS mutant exhibited only slightly increased leucylation activity relative to the T252A single mutation
additional information
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additional information
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leucine-auxotrophic strain
additional information
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construction of an enzyme mutant with a duplication of the peptide fragment from Met238 to Pro368 within the CP1 domain which shows an activity reduced by 59% compared to the wild-type enzyme, and catalyzes the mischarging of tRNALeu by methionine or isoleucine due to impaired ability to edit incorrect products
additional information
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construction of several CP1 domain mutants by introduction of restriction endonuclease sites into gene leuS
additional information
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overexpression of the alpha and beta subunits of the Aquifex aeolicus enzyme and the C- and N-terminal parts of the Escherichia coli enzyme in Escherichia coli as monomeric and dimeric mutants, also mixed between the species, the heterodimeric mutants and the monomeric mutants containing the N-terminal-subpart of the enzyme show very low activity
additional information
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deletion of the unique inserted leucine-specific domain of LeuRS primarily impacts kcat, chimeric LeuRS and ValRS mutants restore limited aminoacylation actiVity, overview
additional information
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deletions of the C terminus differentially impact the two functions of the enzyme in splicing and aminoacylation in vivo, overview, a five-amino acid C-terminal deletion of LeuRS, which does not complement a null strain, can form a ternary complex with the bI4 intron and its maturase splicing partner, however, the complex fails to stimulate splicing activity, deletion of the C-terminal domain of LeuRS abolishes aminoacylation of tRNALeu and also amino acid editing of mischarged tRNA molecules, overview
additional information
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isolated LeuRS CP1 domain requires idiosyncratic adaptations to confer editing activity independent of the full-length enzyme, the beta-strands, which link the CP1 domain to the aminoacylation core of LeuRS, are required for editing of mischarged tRNALeu, hydrolytic activity is also enhanced by inclusion of short flexible peptides, called hinges, at the end of both LeuRS beta-strands, overview
K606R
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kcat/Km: 0.98 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type
additional information
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construction of deletion mutant LeuRS-A lacking residues Q281 to D294, and of deletion mutant LeuRS-B, lacking residues S295 to L304. The Km values of the two mutants for leucine and ATP decrease slightly and for tRNA increase slightly. The kcat values for the three substrates decrease
additional information
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deletion of the entire LSD1 abolishes synthetic activity of LeuRS
additional information
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replacement of Giardia lamblia eukarya-specific insertion 1, GlESI, by human eukarya-specific insertion 1, HsESI, results in mutant DELTAESI/DELTAHsESI. The mutation impairs leucine activation, aminoacylation and post-transfer editing functions without changing the editing specificity
K600R
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the mutation leads to altered catalytic efficiency and perturbations to the discrimination of leucine and isoleucine and affects tRNA recognition and aminoacylation, the mutant shows a slight decrease in activity compared to the wild-type enzyme
additional information
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to explore the oncogenic potential of LARS1 over-expression in lung cancer, LARS1 is knocked-down using siRNA. LARS1 knock-down cells show reduced ability to migrate through transwell membrane and to form colonies in both soft agar and culture plate
K598A
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the mutation simultaneously reduces the tRNA-binding strength and aminoacylation and editing capacities of the enzyme's leucine-specific domain
additional information
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temperature-sensitive and leucine-auxotroph mutant leu-5
K100A/Y109A
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site-directed mutagenesis, mutant steady-state leucine activation kinetics compared to the wild-type enzyme, overview
additional information
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the deletion mutant lacking the C-terminal domain, LeuRSDELTA811-967, retains normal editing activity, but has severely reduced aminoacylation activity, deletion of amino acid residues 911-913 of LeuRS enhances the Ile-tRNAIle deacylation activity, without affecting the Ile-tRNALeu deacylation activity, a C-terminally truncated LeuRS can catalyze the first step of the aminoacylation reaction, Leu-AMP formation, but cannot catalyze the second step, transfer of Leu from Leu-AMP to tRNALeu, overview
additional information
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deletion of the entire LSD1 abolishes synthetic activity of LeuRS
K404Y
P26637
mutant is significantly resistant to inhibitor AN-2690. Mutant aminoacylation of yctRNALeu is not different from wild-type ycLeuRS. Growth rate is similar to wild-type. Growth rate is moderately inhibited in medium containing a large excess of norvaline and reduced leucine
additional information
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respiratory deficient mutants. The phenotype is a consequence of a mutation in a nuclear gene coding for mitochondrial leucyl-tRNA synthetase
additional information
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deletions of the C terminus differentially impact the two functions of the enzyme in splicing and aminoacylation in vivo, overview, a five-amino acid C-terminal deletion of LeuRS, which does not complement a null strain, can form a ternary complex with the bI4 intron and its maturase splicing partner, however, the complex fails to stimulate splicing activity, deletion of the entire yeast mitochondrial LeuRS C-terminal domain enhances its aminoacylation and amino acid editing activities
additional information
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a series of deletions and chimeric variations in the peptide linker of the yeast mitochondrial LeuRS chimeric mutant that is fused to the Escherichia coli LeuRS C-terminal domain extension are created: a four residue deletion mutant of the yeast mitochondrial LeuRS chimera (Ym EcCTD Delta4) stimulates aminoacylation activity significantly compared to that of the chimera enzyme with no deletion within the linker peptide, an eight residue peptide linker deletion mutant that contains three (Ym EcCTD DELTA8/+3), six (Ym EcCTD DELTA8/+6), or nine (Ym EcCTD DELTA8/+9) residues from the Escherichia coli LeuRS linker peptide restors protein stability and activity. Within these three chimeric peptide linker swaps, successive increases in the length of the Escherichia coli chimeric peptide linker decreases aminoacylation activity progressively
additional information
P26637
deletion of the leuS gene is lethal. Human cytoplasmic LeuRS can rescue the knock-out strain but not Escherichia coli LeuRS. LeuRS mutant strains T319, D419A, K404Y, S416D, D418R, T347A or T410A are also able to rescue the null mutant, growth rate of wild-type strain in media containing a large excess of noncognate amino acid norvaline and reduced leucine is reduced to 35%
D347A
Q72GM3
mutation of the highly conserved Asp residue, located in the CP1 domain, is responsible for editing mechanism, slightly reduced activity with L-leucine, mutant mischarges tRNALeu with isoleucine
additional information
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deletion of the C-terminal domain lowering the kcat by 152fold
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
renaturation and refolding of recombinant leuS gene product expressed in Escherichia coli
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
medicine
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LARS1 plays roles in migration and growth of lung cancer cells, which suggest its potential implication in lung tumorigenesis
medicine
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suppression of a tRNA gene mutation by increasing the steady-state levels of its cognate aminoacyl-tRNA synthetase is a model for potential therapies for human pathogenic tRNA mutations
medicine
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the A3243G mutation of the tRNALeu gene causes mitochondrial encephalomyopathy, lactic acidosis, and stroke-like symptoms and 2% of cases of type 2 diabetes. The alteration of aminoacylation of tRNALeu(UUR) caused by the A3243G mutation leads to mitochondrial translational defects and thereby reduces the aminoacylating efficiencies of tRNALeu(UUR) as well as of tRNAAla and tRNAMet. The transfer of human mitochondrial leucyl-tRNA synthetase into the cybrid cells carrying the A3243G mutation improve the efficiency of aminoacylation and stability of mitochondrial tRNAs and then increase the rates of mitochondrial translation and respiration, consequently correcting the mitochondrial dysfunction
medicine
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pretreatment of mice with zinc sulfate solution favors to protection of protein synthesis and acceptor activity of tRNALeu against cadmium-induced inhibition. Under co-exposure of mouse liver to cadmium and zinc, activity of the leucyl-tRNA synthetase is at the level of control. Zinc does not influence TUNEL-positive cell number in cadmium-exposed mouse liver
drug development
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LeuRS is a potential target for antiparasitic drug development