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Literature summary for 6.1.1.4 extracted from

  • 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 (2008), J. Biol. Chem., 283, 22591-22600.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli as a His-tagged fusion protein Homo sapiens
expressed in Escherichia coli as a His-tagged fusion protein Saccharomyces cerevisiae

Protein Variants

Protein Variants Comment Organism
D399K mutant is resitant to inhibitor 5-fluoro-2,1-benzoxaborol-1(3H)-ol but more sensitive to norvaline inhibition Homo sapiens
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 Saccharomyces cerevisiae
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 Saccharomyces cerevisiae
DELTA270-530 deletion of the CP1 domain shows that the mutant is not able to rescue LeuRS knock-out strain Saccharomyces cerevisiae
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 Saccharomyces cerevisiae
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 Saccharomyces cerevisiae
additional information 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 Saccharomyces cerevisiae
additional information growth rate of wild-type strain in media containing a large excess of noncognate amino acid norvaline and reduced leucine is reduced to 35% Saccharomyces cerevisiae
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 Saccharomyces cerevisiae
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 Saccharomyces cerevisiae
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 Saccharomyces cerevisiae
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 Saccharomyces cerevisiae

Inhibitors

Inhibitors Comment Organism Structure
5-fluoro-2,1-benzoxaborol-1(3H)-ol AN-2690, antibiotic which specifically targets the editing active site of LeuRS Homo sapiens
5-fluoro-2,1-benzoxaborol-1(3H)-ol AN-2690, antibiotic which specifically targets the editing active site of LeuRS Saccharomyces cerevisiae
norvaline
-
Homo sapiens

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0041
-
L-leucine
-
Saccharomyces cerevisiae
0.167
-
norvaline
-
Saccharomyces cerevisiae
8.713
-
L-isoleucine
-
Saccharomyces cerevisiae

Localization

Localization Comment Organism GeneOntology No. Textmining
cytoplasm
-
Homo sapiens 5737
-
cytoplasm
-
Saccharomyces cerevisiae 5737
-

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-
Saccharomyces cerevisiae P26637
-
-

Purification (Commentary)

Purification (Comment) Organism
using Ni-NTA chromatography Homo sapiens
using Ni-NTA chromatography Saccharomyces cerevisiae

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-isoleucine + tRNALeu
-
Homo sapiens AMP + diphosphate + L-isoleucyl-tRNALeu
-
?
ATP + L-isoleucine + tRNALeu
-
Saccharomyces cerevisiae AMP + diphosphate + L-isoleucyl-tRNALeu
-
?
ATP + L-leucine + tRNALeu
-
Saccharomyces cerevisiae AMP + diphosphate + L-leucyl-tRNALeu
-
?
ATP + L-norvaline + tRNALeu
-
Saccharomyces cerevisiae AMP + diphosphate + L-norvalyl-tRNALeu
-
?

Synonyms

Synonyms Comment Organism
HcleuRS
-
Homo sapiens
Leucyl-tRNA synthetase
-
Homo sapiens
leucyl—tRNA synthetase
-
Saccharomyces cerevisiae
LeuRS
-
Homo sapiens
LeuRS
-
Saccharomyces cerevisiae
ycLeuRS
-
Saccharomyces cerevisiae

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
30
-
assay at Homo sapiens
30
-
assay at Saccharomyces cerevisiae

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
10.2
-
L-isoleucine
-
Saccharomyces cerevisiae
13.5
-
norvaline
-
Saccharomyces cerevisiae
37.5
-
L-leucine
-
Saccharomyces cerevisiae

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.5
-
assay at Homo sapiens
7.5
-
assay at Saccharomyces cerevisiae