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Information on EC 6.1.1.4 - leucine-tRNA ligase and Organism(s) Saccharomyces cerevisiae and UniProt Accession P26637

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Saccharomyces cerevisiae
UNIPROT: P26637 not found.
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Word Map
The taxonomic range for the selected organisms is: Saccharomyces cerevisiae
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
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
leucyl—tRNA synthetase
-
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
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + L-leucine + tRNALeu = AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
esterification
Aminoacylation
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
L-leucine:tRNALeu ligase (AMP-forming)
-
CAS REGISTRY NUMBER
COMMENTARY hide
9031-15-6
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + L-isoleucine + tRNALeu
AMP + diphosphate + L-isoleucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
-
-
-
?
ATP + L-norvaline + tRNALeu
AMP + diphosphate + L-norvalyl-tRNALeu
show the reaction diagram
-
-
-
?
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-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
ATP + L-methionine + tRNALeu
AMP + diphosphate + L-methionyl-tRNALeu
show the reaction diagram
-
mutant D419A, not the wild-type enzyme
-
r
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
ATP + L-leucine + tRNALeu
AMP + diphosphate + L-leucyl-tRNALeu
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
Mn2+
-
Mg2+, Mn2+, Ca2+ and Ba2+ similarly effective, optimal concentration: 0.1 mM
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5-fluoro-2,1-benzoxaborol-1(3H)-ol
AN-2690, antibiotic which specifically targets the editing active site of LeuRS
Cd2+
-
in presence of 0.1 mM Mg2+
Nalidixic acid
-
-
novobiocin
-
-
Oxolinic acid
-
-
p-chloromercuribenzoate
-
-
Zn2+
-
in presence of 0.1 mM Mg2+
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
8.713
L-isoleucine
-
0.11 - 0.3
ATP
0.02
L-Leu
-
-
0.0001 - 0.0007
tRNALeu
additional information
additional information
-
kinetics of recombinant wild-type and mutant enzymes
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
10.2
L-isoleucine
-
0.05 - 5
tRNALeu
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
assay carried out at room temperature
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
physiological function
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
101900
-
calculation from nucleotide sequence
119000 - 120000
-
equilibrium sedimentation method
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
?
-
x * 55-60000, SDS-PAGE
additional information
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D418R
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
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
deletion of the CP1 domain shows that the mutant is not able to rescue LeuRS knock-out strain
DELTA314-319
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
K404Y
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
S416D
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
T31E
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
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
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
D357A
-
site-directed mutagenesis, the mutant shows reduced activity and abolished editing activity and misaminoacylated isoleucine to tRNALeu compared to the wild-type enzyme
D419A
-
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
DELTA819-828
-
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
-
site-directed mutagenesis, the mutant shows reduced activity and abolished post-transfer editing activity compared to the wild-type enzyme
R449A
-
site-directed mutagenesis, nearly inactive mutant
R449E
-
site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
R449K
-
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
-
site-directed mutagenesis, nearly inactive mutant
R451E
-
site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
R451K
-
site-directed mutagenesis, mutation within the RDW peptide, complementation of the null mutant strain QBY320, 11fold reduced activity compared to the wild-type enzyme
T263V/T264V
-
site-directed mutagenesis, the mutant shows reduced activity and decreased post-transfer editing activity compared to the wild-type enzyme
W445A
-
site-directed mutagenesis, nearly inactive mutant
W445F
-
site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445H
-
site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445K
-
site-directed mutagenesis, mutation within the RDW peptide, no complementation of the null mutant strain QBY320
W445Y
-
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
additional information
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
-
rapid inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
DTT stabilizes
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
using Ni-NTA 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
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expressed in Escherichia coli as a His-tagged fusion protein
expressed in Escherichia coli as a His-tagged fusion protein
-
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 Escherichia coli strain BL21(DE3), complementation abilities of wild-type and mutant enzymes of yeast null strain HM402 and Escherichia coli strain KL321, overview
-
gene CDC60, expression of wild-type and mutants in an Escherichia coli BL21 strain
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Wright, H.T.; Nurse, K.C.; Goldstein, D.J.
Naldixic acid, oxolinic acid, and novobiocin inhibit yeast glycyl- and leucyl-transfer RNA synthetases
Science
213
455-456
1981
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Chirikjian, J.G.; Kanagalingam, K.; Lau, E.; Fresco, J.R.
Purification and properties of leucyl-tRNA synthetase from bakers' yeast
J. Biol. Chem.
248
1074-1079
1973
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Lin, C.S.; Irwin, R.; Chirikjian, J.G.
Kinetic studies of leucyl transfer RNA synthetase from bakers' yeast. Order of addition of substrates and release of products
J. Biol. Chem.
250
9299-9303
1975
Saccharomyces cerevisiae
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
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)
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
Karkhanis, V.A.; Boniecki, M.T.; Poruri, K.; Martinis, S.A.
A viable amino acid editing activity in the leucyl-tRNA synthetase CP1-splicing domain is not required in the yeast mitochondria
J. Biol. Chem.
281
33217-33225
2006
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Nawaz, M.H.; Pang, Y.L.; Martinis, S.A.
Molecular and functional dissection of a putative RNA-binding region in yeast mitochondrial leucyl-tRNA synthetase
J. Mol. Biol.
367
384-394
2007
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Yao, P.; Zhou, X.L.; He, R.; Xue, M.Q.; Zheng, Y.G.; Wang, Y.F.; Wang, E.D.
Unique residues crucial for optimal editing in yeast cytoplasmic Leucyl-tRNA synthetase are revealed by using a novel knockout yeast strain
J. Biol. Chem.
283
22591-22600
2008
Homo sapiens, Saccharomyces cerevisiae (P26637), Saccharomyces cerevisiae
Manually annotated by BRENDA team
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
Manually annotated by BRENDA team
Boniecki, M.T.; Rho, S.B.; Tukalo, M.; Hsu, J.L.; Romero, E.P.; Martinis, S.A.
Leucyl-tRNA synthetase-dependent and -independent activation of a group I intron
J. Biol. Chem.
284
26243-26250
2009
Saccharomyces cerevisiae
Manually annotated by BRENDA team
Duran, R.V.; Hall, M.N.
Leucyl-tRNA synthetase: double duty in amino acid sensing
Cell Res.
22
1207-1209
2012
Saccharomyces cerevisiae, Homo sapiens
Manually annotated by BRENDA team