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Information on EC 6.1.1.4 - leucine-tRNA ligase and Organism(s) Escherichia coli and UniProt Accession P07813
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6.1.1.4 leucine-tRNA ligaseSpecify your search results
Word Map
- 6.1.1.4
- aminoacyl-trna
- synthetases
- aminoacylation
-
fidelity
-
leucylation
-
aarss
-
mischarged
-
post-transfer
- norvaline
-
perrault
-
anticodon
- aeolicus
-
isoacceptors
-
misactivated
-
aquifex
- ilers
-
noncognate
- valrs
-
benzoxaborole
-
isoleucyl-trna
-
misaminoacylated
- hsd17b4
-
kmsks
- metrs
-
pour
- trnaser
-
clinique
- trnaleuuur
- valyl-trna
-
isoleucyl
-
trna-dependent
-
pre-transfer
- drug development
- medicine
The taxonomic range for the selected organisms is: Escherichia coli
The enzyme appears in selected viruses and cellular organisms
The enzyme appears in selected viruses and cellular organisms
Synonyms
leurs, leucyl-trna synthetase, lars2, lars1, ecleurs, cytoplasmic leurs, alphabeta-leurs, glleurs, hs mt leurs, leucyl-trna synthetase 1, 
more
topSYNONYM 
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
Leucine translase
-
-
-
-
Leucine translase
-
-
Leucine--tRNA ligase
-
-
-
-
Leucine--tRNA ligase
-
-
Leucyl-transfer ribonucleate synthetase
-
-
-
-
Leucyl-transfer ribonucleate synthetase
-
-
Leucyl-transfer ribonucleic acid synthetase
-
-
-
-
Leucyl-transfer ribonucleic acid synthetase
-
-
Leucyl-transfer RNA synthetase
-
-
-
-
Leucyl-transfer RNA synthetase
-
-
Leucyl-tRNA synthetase
-
-
-
-
Leucyl-tRNA synthetase
-
-
LeuRS
-
-
-
-
LeuRS
-
-
Synthetase, leucyl-transfer ribonucleate
-
-
-
-
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
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
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
Asp345 is involved in the editing process, mechanism of hydrolysis
-
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
residue E292 is important for aminoacylation activity
-
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
esterification
Aminoacylation
esterification
-
-
-
-
Aminoacylation
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
L-leucine:tRNALeu ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY
9031-15-6
-
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + L-leucine + tRNAGAGLeu
AMP + diphosphate + L-leucyl-tRNAGAGLeu
Escherichia coli tRNAGAGLeu (Ect-RNAGAGLeu)
-
-
?
ATP + L-leucine + tRNAUAALeu
AMP + diphosphate + L-leucyl-tRNAUAALeu
Mycoplasma mobile MmtRNAUAALeu (Mmt-RNAUAALeu)
-
-
?
3'-dATP + L-leucine + tRNALeu
3'-dAMP + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
8-azaadenosine 5'-triphosphate + L-leucine + tRNALeu
8-azaadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
8-bromoadenosine 5'-triphosphate + L-leucine + tRNALeu
8-bromoadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
8-methylaminoadenosine 5'-triphosphate + L-leucine + tRNALeu
8-methylaminoadenosine 5'-monophosphate + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
Adenosine 5'-O-(3-thio)triphosphate + L-leucine + tRNALeu
adenosine 5'-monophosphate + thiodiphosphate + L-leucyl-tRNALeu
-
-
-
-
?
Adenylyl beta,gamma-imido diphosphonate + L-leucine + tRNALeu
Adenylic acid + imido-diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
ATP + 2-butynylalanine + tRNALeu
AMP + diphosphate + 2-butynylalanyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + allylglycine + tRNALeu
AMP + diphosphate + allylglycyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + homoallylglycine + tRNALeu
AMP + diphosphate + homoallylglycyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + homopropargylglycine + tRNALeu
AMP + diphosphate + homopropargylglycyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + L-didehydroleucine + tRNALeu
AMP + diphosphate + didehydroleucyl-tRNALeu
-
reaction is catalyzed by mutant T252Y, not by wild-type
-
-
?
ATP + L-leucine + tRNALeu(UAA)
AMP + diphosphate + L-leucyl-tRNALeu(UAA)
-
-
-
-
?
ATP + L-leucine + tRNALeuCUN
AMP + diphosphate + L-leucyl-tRNALeuCUN
-
-
-
-
?
ATP + L-leucine + tRNALeuUUR
AMP + diphosphate + L-leucyl-tRNALeuUUR
-
-
-
-
?
ATP + L-norisoleucine + tRNALeu
AMP + diphosphate + L-norisoleucyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + L-oxonorvaline + tRNALeu
AMP + diphosphate + oxonorvalyl-tRNALeu
-
reaction is catalyzed by mutant T252Y, not by wild-type
-
-
?
tubercidin 5'-triphosphate + L-leucine + tRNALeu
tubercidin 5'-phosphate + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
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
substrate is tRNALeuTAA, overexpressed in and purified from Escherichia coli
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
-
-
-
-
?
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
-
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
-
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
-
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
-
the ratio of turnover number to KM-value for L-leucine is 1600fold higher than the ratio observed for L-isoleucine
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
mutant T252A edits correctly charged Leu-tRNALeu
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
the connecting peptide CP1 domain is crucial for the editing function
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
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
-
tRNALeu substrates from Escherichia coli and Aquifex aeolicus
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
two-step reaction, the connecting peptide CP1 domain is crucial for the editing function
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
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
-
two functions of the enzyme in splicing and aminoacylation in vivo, overview
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
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
-
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
-
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
-
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-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
-
mutant D345A, not the wild-type enzyme
-
r
ATP + L-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
-
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
-
?
additional information
?
-
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
-
-
?
additional information
?
-
kinetic origin of substrate specificity in post-transfer editing by leucyl-tRNA synthetase, overview. Binding and catalysis is analyzed independently using cognate leucyl- and non-cognate norvalyl-tRNALeu and their non-hydrolyzable analogues. The amino acid part (leucine versus norvaline) of (mis)aminoacyl-tRNAs can contribute approximately 10fold to ground-state discrimination at the editing site, while the rate of deacylation of leucyl- and norvalyl-tRNALeu differs by about 104fold. Critical role for the A76 3'-OH group of the tRNALeu in post-transfer editing. Molecular dynamics simulations reveals that the wild-type enzyme, but not the T252A mutant, enforces leucine to adopt the side-chain conformation that promotes the steric exclusion of a putative catalytic water. Editing can be distiguished from the synthetic site, which relies on ground-state discrimination in amino acid selection
-
-
?
additional information
?
-
-
kinetic origin of substrate specificity in post-transfer editing by leucyl-tRNA synthetase, overview. Binding and catalysis is analyzed independently using cognate leucyl- and non-cognate norvalyl-tRNALeu and their non-hydrolyzable analogues. The amino acid part (leucine versus norvaline) of (mis)aminoacyl-tRNAs can contribute approximately 10fold to ground-state discrimination at the editing site, while the rate of deacylation of leucyl- and norvalyl-tRNALeu differs by about 104fold. Critical role for the A76 3'-OH group of the tRNALeu in post-transfer editing. Molecular dynamics simulations reveals that the wild-type enzyme, but not the T252A mutant, enforces leucine to adopt the side-chain conformation that promotes the steric exclusion of a putative catalytic water. Editing can be distiguished from the synthetic site, which relies on ground-state discrimination in amino acid selection
-
-
?
additional information
?
-
-
leucine-dependent ATP-diphosphate exchange, leucine + ATP + enzyme/Ile-AMP-enzyme + diphosphate
-
-
?
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
?
-
-
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
?
-
-
fidelity of translation is dependent on the specificity of the aminoacyl-tRNA synthetases
-
?
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
?
-
-
isolated LeuRS CP1 domain requires idiosyncratic adaptations to confer editing activity independent of the full-length enzyme, overview
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
ATP + 2-butynylalanine + tRNALeu
AMP + diphosphate + 2-butynylalanyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + allylglycine + tRNALeu
AMP + diphosphate + allylglycyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + homoallylglycine + tRNALeu
AMP + diphosphate + homoallylglycyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + homopropargylglycine + tRNALeu
AMP + diphosphate + homopropargylglycyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + L-norisoleucine + tRNALeu
AMP + diphosphate + L-norisoleucyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
-
aminoacylation by mutant T252Y
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
-
-
r
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
-
two functions of the enzyme in splicing and aminoacylation in vivo, overview
-
-
?
additional information
?
-
the enzyme has evolved both tRNA-dependent pre- and post-transfer editing capabilities to ensure catalytic specificity
-
-
?
additional information
?
-
-
fidelity of translation is dependent on the specificity of the aminoacyl-tRNA synthetases
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
ATP
-
-
ATP
-
optimal concentration for the wild-type enzyme is 4 mM, for the mutants 2 mM
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
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
Mg2+
Zn2+
required, zinc is the molecular switch that controls the catalytic cycle of bacterial leucyl-tRNA synthetase. A specialized zinc-binding domain 1 (ZN-1), when associated with Zn2+, assumes a rigid architecture that is stabilized by thiol groups from the residues C159, C176, and C179. When LeuRS is in the aminoacylation complex, these cysteine residues forman equilateral planar triangular configuration with Zn2+, but when LeuRS transitions to the editing conformation, this geometric configuration breaks down. The wild-type enzyme binds Zn2+ to a greater extent than any of the mutant LeuRSs
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
5-fluoro-1,3-dihydro-1-hydroxy-2,1-benzoxaborole
-
i.e. AN2690, 0.1 mM, 5fold decrease in aminoacylation activity
ATP
-
at high concentration, the mutants are more sensisitve than the wild-type enzyme
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.039
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
0.002
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
0.0021
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
0.0024
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
0.0025
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
2.8
L-isoleucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C
3.5
L-isoleucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C
0.015
L-leucine
-
aminoacylation reaction, wild-type and mutant enzyme, pH 7.8, 37°C
0.052
L-leucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C
0.069
L-leucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C
6.2
L-methionine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C
7.5
L-methionine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C
0.0004
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant wild-type enzyme
-
0.0017
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant mutant L570F
-
0.002
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant mutant L570R
-
0.025
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant mutant L570K
-
0.000018
tRNALeuUUR
-
human derived substrate, pH 7.0, 37°C, recombinant mutant L570K
-
0.00016
tRNALeuUUR
-
human derived substrate, pH 7.0, 37°C, recombinant wild-type enzyme
-
0.0017
tRNALeuUUR
-
human derived substrate, pH 7.0, 37°C, recombinant mutant L570F
-
additional information
additional information
determination of dissociation constants of Zn2+ from LeuRS enzymes
-
additional information
additional information
-
determination of dissociation constants of Zn2+ from LeuRS enzymes
-
additional information
additional information
dissociation constants of LeuRS and mutants from Escherichia coli for their cognate tRNAs. Reaction kinetics of EcLeuRS for Mycoplasma mobile MmtRNAUAALeu and mutant variants, kinetic constants of EcLeuRS, chimeric LeuRS and their mutants for tRNALeu in aminoacylation reaction and for AMP formation in the presence of Nva and MmtRNACAA, detailed overview
-
additional information
additional information
-
dissociation constants of LeuRS and mutants from Escherichia coli for their cognate tRNAs. Reaction kinetics of EcLeuRS for Mycoplasma mobile MmtRNAUAALeu and mutant variants, kinetic constants of EcLeuRS, chimeric LeuRS and their mutants for tRNALeu in aminoacylation reaction and for AMP formation in the presence of Nva and MmtRNACAA, detailed overview
-
additional information
additional information
kinetic origin of substrate specificity in post-transfer editing by leucyl-tRNA synthetase, single-turnover measurements, overview
-
additional information
additional information
-
kinetic origin of substrate specificity in post-transfer editing by leucyl-tRNA synthetase, single-turnover measurements, overview
-
additional information
additional information
-
kinetics, wild-type and mutant enzymes
-
additional information
additional information
-
kinetics of wild-type and mutant enzymes, overview
-
additional information
additional information
-
kinetics of chimeric mutants
-
additional information
additional information
-
kinetics of recombinant His-tagged wild-type and mutant enzymes
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
73.7
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
7
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
7.4
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
7.6
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
8.8
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
6.9
L-isoleucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C
18
L-isoleucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C
101
L-leucine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C
171
L-leucine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C
7.6
L-methionine
-
ATP-diphosphate exchange reaction, mutant enzyme, pH 7.8, 37°C
19
L-methionine
-
ATP-diphosphate exchange reaction, wild-type enzyme, pH 7.8, 37°C
9.8
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant mutant L570K
-
14.5
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant wild-type enzyme
-
21
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant mutant L570R
-
24
tRNALeuCUN
-
Escherichia coli derived substrate, pH 7.0, 37°C, recombinant mutant L570F
-
0.000018
tRNALeuUUR
-
human derived substrate, pH 7.0, 37°C, recombinant mutant L570K
-
0.00016
tRNALeuUUR
-
human derived substrate, pH 7.0, 37°C, recombinant wild-type enzyme
-
0.0017
tRNALeuUUR
-
human derived substrate, pH 7.0, 37°C, recombinant mutant L570F
-
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
1890
L-leucine
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
3100
Ile-tRNALeu
mutant enzyme R286E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
3300
Ile-tRNALeu
wild type enzyme, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
3500
Ile-tRNALeu
mutant enzyme E184R, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
3800
Ile-tRNALeu
mutant enzyme R185E, in 100 mM HEPES (pH 7.8), 10 mM MgCl2, at 37°C
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
0.024
-
mutant A293D, in crude enzyme extract of Escherichia coli strain TG1
0.14
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
1.3
-
purified recombinant wild-type enzyme, with substrate tRNALeu from Aquifex aeolicus
1.5
additional information
0.14
-
mutants A293I, A293G, and A293Y, in crude enzyme extract of Escherichia coli strain TG1
1.5
-
purified recombinant wild-type enzyme, with substrate tRNALeu from Escherichia coli
additional information
-
activiy of several enzyme mutants and mixed mutants, containing subparts of the Escherichia coli enzyme
additional information
-
aminoacid activation activities of wild-type and mutant enzymes
additional information
-
hydrolytic post-transfer editing activity of wild-type and D345A mutant LeuRS CP1 domains, activities of truncation mutants, overview
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
7.5
7.8
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
37
37
-
assay at
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY
GeneOntology No.
LITERATURE
SOURCE
-
during restricted growth on low levels of leucine, localized in cytoplasmic membrane
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
evolution
malfunction
physiological function
additional information
evolution
based on sequence homology and the structures of the catalytic active sites, aaRSs are divided into two classes of 10 members each. Class I synthetases are further divided into three subclasses, a, b, and c, according to sequence homology. Leucyl-tRNA synthetase (LeuRS) belongs to class I aaRSs that include a typical Rossmann dinucleotide-bindingfold active site architecture with the signature sequence modules HIGH and KMSKS. According to evolutionary models, the primitive catalytic core is extended by the insertion and/or fusion of additional domains (also called modules) in LeuRSs, most of which have inserted a large connective polypeptide 1 (CP1) domain that is responsible for amino acid editing. To ensure translation accuracy, LeuRSs have evolved a mechanism to remove aminoacyl AMP (aa-AMP, pre-transfer editing) and aa-tRNA (post-transfer editing). Sequence comparison of the EcLeuRS stem contact-fold domain (SC-fold) with editing-deficient enzymes suggests that key residues of this module have evolved an adaptive strategy to follow the editing functions of LeuRS, basic residues on helix alpha3 of the SC-fold are critical for catalytic efficiency, overview
evolution
enzyme leucyl-tRNA synthetase is part of the aminoacyl-tRNA synthetase (aaRS) family
malfunction
abrogation of the LeuRS specificity determinant threonine 252 does not improve the affinity of the editing site for the cognate leucine as expected, but instead substantially enhances the rate of leucyl-tRNALeu hydrolysis. Molecular dynamics simulations reveals that the wild-type enzyme, but not the T252A mutant, enforces leucine to adopt the side-chain conformation that promotes the steric exclusion of a putative catalytic water
malfunction
mutation of highly conserved basic residues on the third alpha-helix of the KMSKS catalytic loop domain impairs the affinity of LeuRS for the anticodon stem of tRNALeu, which decreases both aminoacylation and editing activities
physiological function
aminoacyl-tRNA synthetases (aaRSs) are a large and diverse family of enzymes that catalyze the attachment of amino acids to their cognate tRNAs in a two-step aminoacylation reaction as follows: 1. amino acid activation by ATP hydrolysis to form an aminoacyl-adenylate intermediate, and 2. transfer of the aminoacyl moiety from the intermediate to the cognate tRNA isoacceptor to form aminoacyl-tRNA (aa-tRNA)
physiological function
Escherichia coli leucyl-tRNA synthetase (LeuRS) is an essential multi-domain metalloenzyme that aminoacylates tRNALeu with leucine. Enzyme LeuRS is an essential enzyme that relies on specialized domains to facilitate the aminoacylation reaction. Structural changes within the ZN-1 domain play a central role in LeuRS's catalytic cycle. The enzyme performs a Zn2+ dependent translocation mechanism for charged tRNALeu, Zn2+ is an architectural cornerstone of the ZN-1 domain and that without its geometric coordination the domain collapses. Residues C159, C176 and C179 coordinate Zn2+ and that this interaction is essential for leucylation to occur, but is not essential for deacylation
physiological function
the ligation of amino acid to tRNA for purposes of protein synthesis proceeds in two steps, bothcatalyzed by a corresponding aminoacyl-tRNA synthetase(aaRS). The amino acid is first activated to anaminoacyl-adenylate (aa-AMP) intermediate at theexpense of ATP, followed by the transfer of aminoacylmoiety to the 2'- or 3'-OH groups at the terminal ribose of the cognate tRNA. Both steps occurwithin the same synthetic/aminoacylation active site located in thecatalytic aaRS domain. Based on the topology of the catalytic domains, the conserved recognition peptides and interaction with the tRNA, aaRSs can be divided into two classes, I and II. The mechanisms of aminoacylation and editing are basically conserved among the classes, although some class-specific features have been recognized. Editing aaRSs exercise specificity through a double-selection mechanism that uses structural/chemical differences between the cognate and non-cognate amino acids twice but in different ways. Leu-tRNALeu is excluded from proofreading basically at the level of catalysis, not binding. This is accomplished by the side chain of the cognate leucine, which adopts a conformation that sterically precludes the positioning of a water nucleophile near the tRNA-assisted hydrolytic machinery. The A76 3'-OH group is a crucial residue in the positioning and activation of the catalytic water. Deacylation mechanism of the enzyme, simulation and modeling, overview
additional information
in silico models of the wild-type and mutated LeuRS CP1 editing domain bound to the analogues with an ester linkage between the amino acid and adenosine as in real substrates [2'-L-leucyladenosine (Leu2A) and 2?-L-norvalyladenosine (Nva2A)] are constructed based on the structure of T252A LeuRS in a complex with tRNALeu and leucyl-adenylate sulphamoyl analogue (Leu-AMS), both positioned in the synthetic active site, and Leu2AA located in the editing domain. The tRNA body dominates the binding energetics of aa-tRNA:LeuRS complex formation
additional information
-
in silico models of the wild-type and mutated LeuRS CP1 editing domain bound to the analogues with an ester linkage between the amino acid and adenosine as in real substrates [2'-L-leucyladenosine (Leu2A) and 2?-L-norvalyladenosine (Nva2A)] are constructed based on the structure of T252A LeuRS in a complex with tRNALeu and leucyl-adenylate sulphamoyl analogue (Leu-AMS), both positioned in the synthetic active site, and Leu2AA located in the editing domain. The tRNA body dominates the binding energetics of aa-tRNA:LeuRS complex formation
additional information
the KMSKS catalytic loop exhibits alpha-alpha-beta-alpha topology in class Ia and Ib aminoacyl-tRNA synthetases, two glycine residues on the third alpha-helix contribute to flexibility, leucine activation, and editing of LeuRS from Escherichia coli (EcLeuRS), acidic residues on the beta-strand enhance the editing activity of EcLeuRS and sense the size of the tRNALeu D-loop. Incorporation of acidic residues on the beta-strand stimulates the tRNA-dependent editing activity of the chimeric minimalist enzyme Mycoplasma mobile LeuRS fused to the connective polypeptide 1 editing domain and leucine-specific domain from EcLeuRS. Sequence comparison of the EcLeuRS stem contact-fold domain with editing-deficient enzymes suggests that key residues of this module have evolved an adaptive strategy to follow the editing functions of LeuRS. Amino acid residues Arg668 or Arg672 are not involved in the amino acid activation step but rather the second tRNA transfer step
additional information
-
the KMSKS catalytic loop exhibits alpha-alpha-beta-alpha topology in class Ia and Ib aminoacyl-tRNA synthetases, two glycine residues on the third alpha-helix contribute to flexibility, leucine activation, and editing of LeuRS from Escherichia coli (EcLeuRS), acidic residues on the beta-strand enhance the editing activity of EcLeuRS and sense the size of the tRNALeu D-loop. Incorporation of acidic residues on the beta-strand stimulates the tRNA-dependent editing activity of the chimeric minimalist enzyme Mycoplasma mobile LeuRS fused to the connective polypeptide 1 editing domain and leucine-specific domain from EcLeuRS. Sequence comparison of the EcLeuRS stem contact-fold domain with editing-deficient enzymes suggests that key residues of this module have evolved an adaptive strategy to follow the editing functions of LeuRS. Amino acid residues Arg668 or Arg672 are not involved in the amino acid activation step but rather the second tRNA transfer step
additional information
there are two catalytic sites, the leucylation site housed within the aminoacylation domain and the hydrolytic deacylation site housed within the CP1 editing domain, the ZN-1 domain is known to play an essential structural role in stabilizing the Leu-Amp adenylate. Structural analysis of LeuRS enzymes using Fourier transform infrared spectroscopy (FTIR), and homology modeling of LeuRS in the editing conformation, visualizing the ZN-1 domain, by using the structure in editing conformation of the LeuRS enzyme from Thermus thermophilus, PDB ID 1OBH as template, overview
additional information
-
there are two catalytic sites, the leucylation site housed within the aminoacylation domain and the hydrolytic deacylation site housed within the CP1 editing domain, the ZN-1 domain is known to play an essential structural role in stabilizing the Leu-Amp adenylate. Structural analysis of LeuRS enzymes using Fourier transform infrared spectroscopy (FTIR), and homology modeling of LeuRS in the editing conformation, visualizing the ZN-1 domain, by using the structure in editing conformation of the LeuRS enzyme from Thermus thermophilus, PDB ID 1OBH as template, overview
PDB
SCOP
CATH
UNIPROT
ORGANISM
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
34000
-
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
63000
-
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
97300
-
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
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
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
?
-
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
additional information
structure comparison of the Zn-1 domain in editing and in aminoacylation conformations, overview
additional information
-
structure comparison of the Zn-1 domain in editing and in aminoacylation conformations, overview
additional information
-
determination of secondary structure elements in the wild-type and mutant enzyme
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
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
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
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
enzyme LeuRS mutant T252A in a complex with tRNALeu and leucyl-adenylate sulphamoyl analogue (Leu-AMS), both positioned in the synthetic active site, and Leu2AA located in the editing domain, X-ray diffraction structure determination and analysis at resolution, replacement using structure PDB ID 4AQ7, modeling
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
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
C159A
site-directed mutagenesis, structure comparison with the wild-type
C176A
site-directed mutagenesis, structure comparison with the wild-type
C179A
site-directed mutagenesis, structure comparison with the wild-type
D342A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
D345A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
E184A
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
E184R
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
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
K671A
site-directed mutagenesis, the mutation does no affect the catalytic efificiency
R185E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
R286E
the mutation significantly enhances tRNA-dependent pre-transfer editing activity
R344A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
R668A
site-directed mutagenesis, the mutant shows 77% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type
R668A/R672A
site-directed mutagenesis, the mutant shows 93.6% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type
R668E
site-directed mutagenesis, the mutant shows 95% reduced catalytic efficiency compared to wild-type, the rate of AMP formation is decreased compared to the wild-type
R668E/R672E
site-directed mutagenesis, the mutant shows 98.6% reduced catalytic efficiency compared to wild-type. But the almost inactive mutant exhibits intact Leu activation activity comparable with the wild-type enzyme
R672A
site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type
R672E
site-directed mutagenesis, the rate of AMP formation is decreased compared to the wild-type
T248A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, overview
T252A
site-directed mutagenesis, the mutant shows altered deacylation activity with amino acids norvaline, isoleucine, and leucine compared to the wild-type enzyme, conformational changes associated with the binding of post-transfer editing analogues in the editing site of T252A LeuRS, overview
T252R
the mutant performs the activities of amino acid activation, aminoacylation and deacylation of mischarged tRNAs as well as the native enzyme
A293D
A293E
-
site-directed mutagenesis, the mutant activity is similar to the wild-type enzyme
A293F
-
54% decreased activity compared to the wild-type, more sensitive too inhibition by ATP
A293G
-
50% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
A293I
-
51% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates, more sensitive too inhibition by ATP
A293K
-
site-directed mutagenesis, the post-transfer editing activity of the isolated CP1-domain is enhanced compared to the wild-type enzyme's domain
A293R
A293Y
-
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
DELTA788-798
-
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
-
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
-
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
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA794
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA794-798
-
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
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA795-796
-
two-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA795-798
-
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
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA796-798
DELTA797
-
single-site deletion at the more flexible end of the peptide linker: no significant change in aminoacylation activity
DELTA797-798
-
two-site deletion at the more flexible end of the peptide linker: mutant exhibits lower aminoacylation activity compared to wild-type
E292A
-
unaltered specific activity in amino acid activation reaction, 61% reduced aminoacylation activity compared to the wild-type
E292D
-
unaltered specific activity in amino acid activation reaction, 53% reduced aminoacylation activity compared to the wild-type
E292F
-
unaltered specific activity in amino acid activation reaction, 60% reduced aminoacylation activity compared to the wild-type
E292K
-
unaltered specific activity in amino acid activation reaction, 85% reduced aminoacylation activity compared to the wild-type
E292Q
-
unaltered specific activity in amino acid activation reaction, 54% reduced aminoacylation activity compared to the wild-type
E292S
-
unaltered specific activity in amino acid activation reaction, 66% reduced aminoacylation activity compared to the wild-type
E797GGG
-
mutant shows no altered aminoacylation activity compared to wild-type
E797PPP
-
mutant shows no altered aminoacylation activity compared to wild-type
G225P
-
abolishes tRNA leucylation due to a defect in leucine activation, decrease in deacylation of Ile-tRNALeu
G229P
-
increased aminoacylation activity compared to the wild-type, mutant deacylates Ile-tRNALeu similar to wild-type
G229P/T252A
-
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
-
aminoacylates tRNALeu and decylates Ile-tRNALeu as well as wild-type
G409P
-
increased aminoacylation activity compared to the wild-type, mutant deacylates Ile-tRNALeu similar to wild-type
K809A
-
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
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K846E
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
K846E/K853E
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
K853A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-typ enzyme
K853E
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
L570F
-
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
-
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
-
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
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
L855A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
M336A
-
site-directed mutagenesis, editing site mutant, the mutant shows a small increase in leucine editing activity
M336F/T252A
-
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
N807A
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
N807A/N856A
-
site-directed mutagenesis, the mutant shows similar activity compared to the wild-typ enzyme
N856A
-
site-directed mutagenesis, the mutant shows increased activity compared to the wild-typ enzyme
Q805A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
Q805A/N807A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
Q805A/N807A/N856A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
R249F
-
site-directed mutagenesis, editing site mutant, editing activity of Leu-tRNALeu is decreased
R249F/T252A
-
site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity
R249T
-
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
-
site-directed mutagenesis, editing site mutant, the mutant shows decreased hydrolysis of mischarged Ile-tRNALeu compared to the wild type enzyme
R811A
-
site-directed mutagenesis, the mutant shows decreased activity compared to the wild-typ enzyme
T247A/T248A
-
533fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247S/T248S
-
77fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V
T247V/T248V
T248V
T252A
T252D
-
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
T252F
-
impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine
T252G
-
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
-
impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine
T252S
-
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
-
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
T272R
-
no change in aminoacylation activity, but the deacylation of Ile-tRNALeu is strongly impaired. Mutant still exhibits 45% of wild-type AMP formation
V338A
-
site-directed mutagenesis, editing site mutant, it shows increased post-transfer editing activity of Leu-tRNALeu compared to the wild-type enzyme
V338D
-
site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
V338E
-
site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
V338F
-
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
-
site-directed mutagenesis, editing site mutant, the T252A mutation uncouples specificity
V338L
-
site-directed mutagenesis, editing site mutant, the mutant shows reduced post-transfer editing activity compared to the wild-type enzyme
additional information
A293D
-
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
-
site-directed mutagenesis, the mutant activity is similar to the wild-type enzyme
A293R
-
30% decreased activity, decreased editing function, stronger binding of ATP, decrease in Km for the substrates
A293R
-
site-directed mutagenesis, the post-transfer editing activity of the isolated CP1-domain is enhanced compared to the wild-type enzyme's domain
D345A
-
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
DELTA796-798
-
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
DELTA796-798
-
two-site deletion at the more flexible end of the peptide linker: mutant exhibits lower aminoacylation activity compared to wild-type
T247V
-
8fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V
-
site-directed mutagenesis, hydrolysis of Ile-tRNALeu is completely abolished
T247V/T248V
-
fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T247V/T248V
-
site-directed mutagenesis, the double mutation abolishes post-transfer editing activity
T248V
-
6fold decrease in the ratio of turnover number to Km-value compared to wild-type ratio
T248V
-
site-directed mutagenesis, hydrolysis of Ile-tRNALeu is completely abolished
T252A
-
decreased activity with L-leucine, mutant shows altered editing specificity, it edits correctly formed leucyl-tRNALeu
T252A
-
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
-
site-directed mutagenesis, the mutation in the full-length LeuRS uncouples specificity and hydrolyzes correctly charged LeutRNALeu
T252A
-
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
T252E
-
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
T252Y
-
impaired proofreading mechanism, increase rate of misaminoacylation with isoleucine and valine, effective aminoacylation of tRNALeu with allylglycine, homopropargylglycine, 2-butynylalanine, norvaline, and norisoleucine
T252Y
-
site-directed mutagenesis, the mutation occupies the amino acid binding pocket and blocks the binding of substrate to abolish editing activity
T252Y
-
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
additional information
C159A, C176A and C179A disassociate from Zn2+ much more readily than wild-type LeuRS, the wild-type LeuRS binds more tightly to Zn2+ than do C159A, C176A or C179A
additional information
-
C159A, C176A and C179A disassociate from Zn2+ much more readily than wild-type LeuRS, the wild-type LeuRS binds more tightly to Zn2+ than do C159A, C176A or C179A
additional information
in silico models of the wild-type and mutated LeuRS CP1 editing domain bound to the analogues with an ester linkage between the amino acid and adenosine as in real substrates [2'-L-leucyladenosine (Leu2A) and 2?-L-norvalyladenosine (Nva2A)] are constructed based on the structure of T252A LeuRS in a complex with tRNALeu and leucyl-adenylate sulphamoyl analogue (Leu-AMS), both positioned in the synthetic active site, and Leu2AA located in the editing domain. The tRNA body dominates the binding energetics of aa-tRNA:LeuRS complex formation
additional information
-
in silico models of the wild-type and mutated LeuRS CP1 editing domain bound to the analogues with an ester linkage between the amino acid and adenosine as in real substrates [2'-L-leucyladenosine (Leu2A) and 2?-L-norvalyladenosine (Nva2A)] are constructed based on the structure of T252A LeuRS in a complex with tRNALeu and leucyl-adenylate sulphamoyl analogue (Leu-AMS), both positioned in the synthetic active site, and Leu2AA located in the editing domain. The tRNA body dominates the binding energetics of aa-tRNA:LeuRS complex formation
additional information
-
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
-
construction of several CP1 domain mutants by introduction of restriction endonuclease sites into gene leuS
additional information
-
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
-
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
-
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
-
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
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
49.5
-
50% inactivation, heating of 0.5°C per minute starting at 30°C, mutant A293D
54
-
50% inactivation, heating of 0.5°C per minute starting at 30°C, wild-type enzyme and mutants A293F, A293G, A293R
55
-
50% inactivation, heating of 0.5°C per minute starting at 30°C, mutants A293I and A293Y
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20°C, 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
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21 by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography, co-purifying Leu-AMP is removed
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 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 enzyme from overexpression in Escherichia coli, purification of the proteolytically derived fragments from wild-type enzyme preparation
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
gene leuS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21 from plasmid pRWecLeuRS
recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli 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
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)
-
gene leuS, expression of His-tagged wild-type and mutant enzymes in strain BL21(DE3)
-
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
-
subcloning in strain DH5alpha, expression of His-tagged wild-type and mutant enzymes in strain BL21(DE3)
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Williamson, R.M.
Membrane association of leucyl-tRNA synthetase during leucine starvation in Escherichia coli
Biochem. Biophys. Res. Commun.
190
794-800
1993
Escherichia coli
Berg, P.; Bergmann, F.H.; Ofengand, E.J.; Dieckmann, M.
The enzymic synthesis of amino acyl derivatives of ribonucleic acid. The mechanism of leucyl-, valyl-, isoleucyl-, and methionyl ribonucleic acid formation
J. Biol. Chem.
236
1726-1734
1961
Escherichia coli
-
Marutzky, R.; Flossdorf, J.; Kula, M.R.
ATP-analogue as substrates for leucyl-TRNA synthetase from Escherichia coli MRE 600
Nucleic Acids Res.
3
2067-2077
1976
Escherichia coli, Escherichia coli MRE 600
Tzagoloff, A.; Akai, A.; Kurkulos, M.; Repetto, B.
Homology of yeast mitochondrial leucyl-tRNA synthetase and isoleucyl- and methionyl-tRNA synthetases of Escherichia coli
J. Biol. Chem.
263
850-856
1988
Saccharomyces cerevisiae, Escherichia coli
Herbert, C.J.; Labouesse, M.; Dujardin, G.; Slonimski, P.P.
The NAM2 proteins from S. cerevisiae and S. douglasii are mitochondrial leucyl-tRNA synthetases, and are involved in mRNA splicing
EMBO J.
7
473-483
1988
Escherichia coli, Saccharomyces douglasii
Bergmann, F.H.; Berg, P.; Dieckmann, M.
The Enzymic synthesis of amino acyl derivatives of ribonucleic acid. II. The preparation of leucyl-, valyl-, isoleucyl-, and methionyl ribonucleic acid synthetases from Escherichia coli
J. Biol. Chem.
236
1735-1740
1961
Escherichia coli
-
Li, T.; Guo, N.; Xia, X.; Wang, E.D.; Wang, Y.L.
The peptide bond between E292-A293 of Escherichia coli leucyl-tRNA synthetase is essential for its activity
Biochemistry
38
13063-13069
1999
Escherichia coli
Chen, J.F.; Guo, N.N.; Li, T.; Wang, E.D.; Wang, Y.L.
CP1 domain in Escherichia coli leucyl-tRNA synthetase is crucial for its editing function
Biochemistry
39
6726-6731
2000
Escherichia coli
Chen, J.F.; Li, T.; Wang, E.D.; Wang, Y.L.
Effect of alanine-293 replacement on the activity, ATP binding, and editing of Escherichia coli leucyl-tRNA synthetase
Biochemistry
40
1144-1149
2001
Escherichia coli
Tang, Y.; Tirrell, D.A.
Attenuation of the editing activity of the Escherichia coli leucyl-tRNA synthetase allows incorporation of novel amino acids into proteins in vivo
Biochemistry
41
10635-10645
2002
Escherichia coli
Zhao, M.W.; Hao, R.; Chen, J.F.; Martin, F.; Eriani, G.; Wang, E.D.
Enzymes assembled from Aquifex aeolicus and Escherichia coli leucyl-tRNA synthetases
Biochemistry
42
7694-7700
2003
Aquifex aeolicus, Escherichia coli
Du, X.; Wang, E.D.
E292 is important for the aminoacylation activity of Escherichia coli leucyl-tRNA synthetase
J. Protein Chem.
22
71-76
2003
Escherichia coli
Lincecum, T.L., Jr.; Tukalo, M.; Yaremchuk, A.; Mursinna, R.S.; Williams, A.M.; Sproat, B.S.; Van Den Eynde, W.; Link, A.; Van Calenbergh, S.; Grotli, M.; Martinis, S.A.; Cusack, S.
Structural and mechanistic basis of pre- and posttransfer editing by leucyl-tRNA synthetase
Mol. Cell
11
951-963
2003
Saccharomyces cerevisiae, Escherichia coli, Thermus thermophilus (Q72GM3)
Chen, J.; Li, Y.; Wang, E.; Wang, Y.
High-level expression and single-step purification of leucyl-tRNA synthetase from Escherichia coli
Protein Expr. Purif.
15
115-120
1999
Escherichia coli
Xu, M.G.; Li, J.; Du, X.; Wang, E.D.
Groups on the side chain of T252 in Escherichia coli leucyl-tRNA synthetase are important for discrimination of amino acids and cell viability
Biochem. Biophys. Res. Commun.
318
11-16
2004
Escherichia coli
Liu, Y.; Liao, J.; Zhu, B.; Wang, E.; Ding, J.
Crystal structures of the editing domain of E. coli leucyl-tRNA synthetase and its complexes with methionine and isoleucine reveal a lock-and-key mechanism for amino acid discrimination
Biochem. J.
394
399-407
2006
Escherichia coli (P07813), Escherichia coli
Mursinna, R.S.; Lee, K.W.; Briggs, J.M.; Martinis, S.A.
Molecular dissection of a critical specificity determinant within the amino acid editing domain of leucyl-tRNA synthetase
Biochemistry
43
155-165
2004
Escherichia coli
Zhai, Y.; Martinis, S.A.
Two conserved threonines collaborate in the Escherichia coli leucyl-tRNA synthetase amino acid editing mechanism
Biochemistry
44
15437-15443
2005
Escherichia coli
Lue, S.W.; Kelley, S.O.
An aminoacyl-tRNA synthetase with a defunct editing site
Biochemistry
44
3010-3016
2005
Escherichia coli, Homo sapiens
Lee, K.W.; Briggs, J.M.
Molecular modeling study of the editing active site of Escherichia coli leucyl-tRNA synthetase: two amino acid binding sites in the editing domain
Proteins
54
693-704
2004
Escherichia coli
Zhai, Y.; Nawaz, M.H.; Lee, K.W.; Kirkbride, E.; Briggs, J.M.; Martinis, S.A.
Modulation of substrate specificity within the amino acid editing site of leucyl-tRNA synthetase
Biochemistry
46
3331-3337
2007
Escherichia coli
Lue, S.W.; Kelley, S.O.
A single residue in leucyl-tRNA synthetase affecting amino acid specificity and tRNA aminoacylation
Biochemistry
46
4466-4472
2007
Escherichia coli, Homo sapiens
Vu, M.T.; Martinis, S.A.
A unique insert of leucyl-tRNA synthetase is required for aminoacylation and not amino acid editing
Biochemistry
46
5170-5176
2007
Escherichia coli
Betha, A.K.; Williams, A.M.; Martinis, S.A.
Isolated CP1 domain of Escherichia coli leucyl-tRNA synthetase is dependent on flanking hinge motifs for amino acid editing activity
Biochemistry
46
6258-6267
2007
Escherichia coli
Hsu, J.L.; Rho, S.B.; Vannella, K.M.; Martinis, S.A.
Functional divergence of a unique C-terminal domain of leucyl-tRNA synthetase to accommodate its splicing and aminoacylation roles
J. Biol. Chem.
281
23075-23082
2006
Saccharomyces cerevisiae, Escherichia coli
Mascarenhas, A.P.; Martinis, S.A.
Functional segregation of a predicted hinge site within the beta-strand linkers of Escherichia coli leucyl-tRNA synthetase
Biochemistry
47
4808-4816
2008
Escherichia coli
Hsu, J.L.; Martinis, S.A.
A flexible peptide tether controls accessibility of a unique C-terminal RNA-binding domain in Leucyl-tRNA synthetases
J. Mol. Biol.
376
482-491
2008
Saccharomyces cerevisiae, Escherichia coli
Tang, Y.; Wang, P.; Van Deventer, J.; Link, A.; Tirrell, D.
Introduction of an aliphatic ketone into recombinant proteins in a bacterial strain that overexpresses an editing-impaired leucyl-tRNA synthetase
ChemBioChem
10
2188-2190
2009
Escherichia coli
Mascarenhas, A.P.; Martinis, S.A.
A glycine hinge for tRNA-dependent translocation of editing substrates to prevent errors by leucyl-tRNA synthetase
FEBS Lett.
583
3443-3447
2009
Escherichia coli
Tan, M.; Zhu, B.; Zhou, X.L.; He, R.; Chen, X.; Eriani, G.; Wang, E.D.
tRNA-dependent pre-transfer editing by prokaryotic leucyl-tRNA synthetase
J. Biol. Chem.
285
3235-3244
2010
Aquifex aeolicus, Escherichia coli
Tan, M.; Zhu, B.; Liu, R.J.; Chen, X.; Zhou, X.L.; Wang, E.D.
Interdomain communication modulates the tRNA-dependent pre-transfer editing of leucyl-tRNA synthetase
Biochem. J.
449
123-131
2013
Escherichia coli (P07813)
Yan, W.; Ye, Q.; Tan, M.; Chen, X.; Eriani, G.; Wang, E.D.
Modulation of aminoacylation and editing properties of leucyl-tRNA synthetase by a conserved structural module
J. Biol. Chem.
290
12256-12267
2015
Pyrococcus horikoshii (O58698), Aquifex aeolicus (O66680 AND O67646), Escherichia coli (P07813), Escherichia coli, Mesomycoplasma mobile (Q6KHA5), Homo sapiens (Q9P2J5), Mesomycoplasma mobile ATCC 43663 / 163K / NCTC 11711 (Q6KHA5), Pyrococcus horikoshii ATCC 700860 / DSM 12428 / JCM 9974 / NBRC 100139 / OT-3 (O58698)
Kumar, M.; Kumar, S.A.; Dimkovikj, A.; Baykal, L.N.; Banton, M.J.; Outlaw, M.M.; Polivka, K.E.; Hellmann-Whitaker, R.A.
Zinc is the molecular switch that controls the catalytic cycle of bacterial leucyl-tRNA synthetase
J. Inorg. Biochem.
142
59-67
2015
Escherichia coli (P07813), Escherichia coli
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