Cloned (Comment) | Organism |
---|---|
expression in Escherichia coli strain KL231 cells | Giardia intestinalis |
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 | 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 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 | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Giardia intestinalis |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Giardia intestinalis | - |
- |
- |
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 | Comment | Organism |
---|---|---|
Leucyl-tRNA synthetase | - |
Giardia intestinalis |
LeuRS | - |
Giardia intestinalis |
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 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 Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.6 | - |
assay at, with HstRNALeu | Giardia intestinalis |
8.2 | - |
assay at, with GltRNALeu | Giardia intestinalis |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Giardia intestinalis |
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 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 |