Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 6.1.1.4 extracted from

  • Zhou, X.L.; Tan, M.; Wang, M.; Chen, X.; Wang, E.D.
    Post-transfer editing by a eukaryotic leucyl-tRNA synthetase resistant to the broad-spectrum drug AN2690 (2010), Biochem. J., 430, 325-333.
    View publication on PubMed
Search for more literature for 6.1.1.4

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli strain KL231 cells Giardia intestinalis

Protein Variants

Protein Variants Comment Organism
D444A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis
D444E site-directed mutagenesis Giardia intestinalis
additional information 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 Giardia intestinalis
R338A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis
T341A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis
T341R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis

Inhibitors

Inhibitors Comment Organism Structure
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 Giardia intestinalis
additional information development of a GlLeuRS-specific inhibitor for the treatment of giardiasis Giardia intestinalis

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0056
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341A Giardia intestinalis
0.0057
-
 tRNALeu pH 8.2, 45°C, recombinant mutant D444A Giardia intestinalis
0.0059
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341R Giardia intestinalis
0.008
-
 tRNALeu pH 8.2, 45°C, recombinant mutant R338A Giardia intestinalis
0.0093
-
 tRNALeu pH 8.2, 45°C, recombinant mutant DELTAESI/DELTAHsESI Giardia intestinalis

Metals/Ions

Metals/Ions Comment Organism Structure
Mg2+
-
 Giardia intestinalis

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
ATP + L-leucine + tRNALeu Giardia intestinalis
-
 AMP + diphosphate + L-leucyl-tRNALeu
-
 ?

Organism

Organism UniProt Comment Textmining
Giardia intestinalis
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
ATP + L-leucine + tRNALeu
-
 Giardia intestinalis AMP + diphosphate + L-leucyl-tRNALeu
-
 ?
ATP + L-leucine + tRNALeu tRNAs from Homo sapiens and Giardia lamblia Giardia intestinalis AMP + diphosphate + L-leucyl-tRNALeu
-
 ?

Synonyms

Synonyms Comment Organism
Leucyl-tRNA synthetase
-
 Giardia intestinalis
LeuRS
-
 Giardia intestinalis

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
37
-
 assay at, with HstRNALeu Giardia intestinalis
45
-
 assay at, with GltRNALeu Giardia intestinalis

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.7
-
 tRNALeu pH 8.2, 45°C, recombinant mutant DELTAESI/DELTAHsESI Giardia intestinalis
1
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341A Giardia intestinalis
1.1
-
 tRNALeu pH 8.2, 45°C, recombinant mutant D444A Giardia intestinalis
1.1
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341R Giardia intestinalis
1.3
-
 tRNALeu pH 8.2, 45°C, recombinant mutant R338A Giardia intestinalis

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7.6
-
 assay at, with HstRNALeu Giardia intestinalis
8.2
-
 assay at, with GltRNALeu Giardia intestinalis

Cofactor

Cofactor Comment Organism Structure
ATP
-
 Giardia intestinalis

General Information

General Information Comment Organism
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 Giardia intestinalis
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 Giardia intestinalis

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
75.3
-
 tRNALeu pH 8.2, 45°C, recombinant mutant DELTAESI/DELTAHsESI Giardia intestinalis
162.5
-
 tRNALeu pH 8.2, 45°C, recombinant mutant R338A Giardia intestinalis
178.6
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341A Giardia intestinalis
186.4
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341R Giardia intestinalis
193
-
 tRNALeu pH 8.2, 45°C, recombinant mutant D444A Giardia intestinalis