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Literature summary for 6.1.1.4 extracted from
- The CP2 domain of leucyl-tRNA synthetase is crucial for amino acid activation and post-transfer editing (2008), J. Biol. Chem., 283, 36608-36616.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| expressed in Escherichia coli as a His-tagged fusion protein | Giardia intestinalis |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| D588A | kcat/Km: 0.57 (ATP), 0.37 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type | Giardia intestinalis |
| D603A | kcat/Km: 0.91 (ATP), 0.94 (Leu), similar activity compared to wild-type | Giardia intestinalis |
| DELTA581-617 | deletion of the CP2 domain, an insertion domain called connective peptide 2, of Giardia lamblia LeuRS shows that the CP2 domain is indispensable for amino acid activation, post-transfer editing and contributes to LeuRS-tRNALeu binding affinity. CP2 domain of Pyrococcus horikoshii LeuRS but not that of Escherichia coli LeuRS can partially restore amino acid activation and post-transfer editing functions suggesting that the functions of the CP2 domain are dependent on its location in the primary sequence of LeuRS | Giardia intestinalis |
| K587A | kcat/Km: 0.43 (ATP), 0.42 (Leu), mutant displays lower amino acid activation and aminoacylation activities than wild-type | Giardia intestinalis |
| K606A | mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities | Giardia intestinalis |
| K606D | kcat/Km: 0.86 (ATP), 0.83 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type | Giardia intestinalis |
| K606E | kcat/Km: 0.90 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type | Giardia intestinalis |
| K606L | kcat/Km: 0.85 (ATP), 0.98 (Leu), similar leucine activation and post-transfer editing activities compared to wild-type | Giardia intestinalis |
| K606R | kcat/Km: 0.98 (ATP), 0.99 (Leu), no difference in leucine activation and post-transfer editing activities compared to wild-type | Giardia intestinalis |
| W586A | mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities | Giardia intestinalis |
| Y581A | mutant shows a complete loss of amino acid activation, aminoacylation and post-transfer editing activities | Giardia intestinalis |
| Y581E | mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity | Giardia intestinalis |
| Y581F | mutant shows some but significantly reduced leucine activation activity its post-transfer editing activity is similar to wild-type | Giardia intestinalis |
| Y581K | mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity | Giardia intestinalis |
| Y581S | mutant is unable to activate leucine by the ATP-diphosphate exchange assay, and mutant has no post-transfer editing activity | Giardia intestinalis |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kcat/Km: 1 (ATP, wild-type), 1 (Leu, wild-type), 0.43 (ATP, mutant K587A), 0.42 (Leu, mutant K587A), 0.57 (ATP, mutant K588A), 0.37 (Leu, mutant K588A), 0.91 (ATP, mutant D603A), 0.94 (Leu, mutant D603A), 0.98 (ATP, mutant K606R), 0.99 (Leu, mutant K606R), 0.9 (ATP, mutant K606E), 0.99 (Leu, mutant K606E), 0.85 (ATP, mutant K606L), 0.98 (Leu, mutant K606L), 0.86 (ATP, mutant K606D), 0.83 (Leu, mutant K606D) | Giardia intestinalis |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Giardia intestinalis | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| using Ni-NTA chromatography | Giardia intestinalis |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-isoleucine + tRNALeu | - |
Giardia intestinalis | AMP + diphosphate + L-isoleucyl-tRNALeu | - |
? |  |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| GlLeuRS | - |
Giardia intestinalis |
| Leucyl-tRNA synthetase | - |
Giardia intestinalis |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | - |
assay at | Giardia intestinalis |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 8.2 | - |
assay at | Giardia intestinalis |
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