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Literature summary 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

Cloned(Commentary)

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

Protein Variants

EC Number Protein Variants Comment Organism
6.1.1.4 D444A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis
6.1.1.4 D444E site-directed mutagenesis Giardia intestinalis
6.1.1.4 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
6.1.1.4 R338A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis
6.1.1.4 T341A site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis
6.1.1.4 T341R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Giardia intestinalis

Inhibitors

EC Number Inhibitors Comment Organism Structure
6.1.1.4 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
6.1.1.4 additional information development of a GlLeuRS-specific inhibitor for the treatment of giardiasis Giardia intestinalis

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
6.1.1.4 0.0056
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341A Giardia intestinalis
6.1.1.4 0.0057
-
 tRNALeu pH 8.2, 45°C, recombinant mutant D444A Giardia intestinalis
6.1.1.4 0.0059
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341R Giardia intestinalis
6.1.1.4 0.008
-
 tRNALeu pH 8.2, 45°C, recombinant mutant R338A Giardia intestinalis
6.1.1.4 0.0093
-
 tRNALeu pH 8.2, 45°C, recombinant mutant DELTAESI/DELTAHsESI Giardia intestinalis

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
6.1.1.4 Mg2+
-
 Giardia intestinalis

Natural Substrates/ Products (Substrates)

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

Organism

EC Number Organism UniProt Comment Textmining
6.1.1.4 Giardia intestinalis
-
-
-

Substrates and Products (Substrate)

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

Synonyms

EC Number Synonyms Comment Organism
6.1.1.4 Leucyl-tRNA synthetase
-
 Giardia intestinalis
6.1.1.4 LeuRS
-
 Giardia intestinalis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
6.1.1.4 37
-
 assay at, with HstRNALeu Giardia intestinalis
6.1.1.4 45
-
 assay at, with GltRNALeu Giardia intestinalis

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
6.1.1.4 0.7
-
 tRNALeu pH 8.2, 45°C, recombinant mutant DELTAESI/DELTAHsESI Giardia intestinalis
6.1.1.4 1
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341A Giardia intestinalis
6.1.1.4 1.1
-
 tRNALeu pH 8.2, 45°C, recombinant mutant D444A Giardia intestinalis
6.1.1.4 1.1
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341R Giardia intestinalis
6.1.1.4 1.3
-
 tRNALeu pH 8.2, 45°C, recombinant mutant R338A Giardia intestinalis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
6.1.1.4 7.6
-
 assay at, with HstRNALeu Giardia intestinalis
6.1.1.4 8.2
-
 assay at, with GltRNALeu Giardia intestinalis

Cofactor

EC Number Cofactor Comment Organism Structure
6.1.1.4 ATP
-
 Giardia intestinalis

General Information

EC Number General Information Comment Organism
6.1.1.4 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
6.1.1.4 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]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
6.1.1.4 75.3
-
 tRNALeu pH 8.2, 45°C, recombinant mutant DELTAESI/DELTAHsESI Giardia intestinalis
6.1.1.4 162.5
-
 tRNALeu pH 8.2, 45°C, recombinant mutant R338A Giardia intestinalis
6.1.1.4 178.6
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341A Giardia intestinalis
6.1.1.4 186.4
-
 tRNALeu pH 8.2, 45°C, recombinant mutant T341R Giardia intestinalis
6.1.1.4 193
-
 tRNALeu pH 8.2, 45°C, recombinant mutant D444A Giardia intestinalis