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Literature summary for 6.1.1.4 extracted from
- Split leucine-specific domain of leucyl-tRNA synthetase from the hyperthermophilic bacterium Aquifex aeolicus (2006), Biochemistry, 45, 14809-14816.
Cloned(Commentary)
| Cloned (Comment) | Organism |
|---|---|
| individual expression of His-tagged alpha- and beta-subunits in Escherichia coli strain BL21(DE3) | Aquifex aeolicus |
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| K599A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| K599A/K600A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| K599A/K600A/K605A/K606A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| K600A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| K605A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| K605A/K606A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| K606A | site-directed mutagenesis, the mutation of the residue in the alpha-subunit involved in catalysis only slightly affects the enzyme activity | Aquifex aeolicus |
| additional information | construction of diverse truncation mutants, ATP-diphosphate-exchange and aminoacylation activities, and ability to charge minihelixLeu of alpha and beta subunit mutants, overview | Aquifex aeolicus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| additional information | - |
additional information | kinetics of recombinant trunacted mutants, overview | Aquifex aeolicus | |
| 0.0003 | - |
tRNALeuGAG | pH 6.8, 65°C, recombinant wild-type enzyme | Aquifex aeolicus | |
| 0.006 | - |
L-leucine | pH 6.8, 65°C, recombinant wild-type enzyme | Aquifex aeolicus |  |
| 0.1123 | - |
ATP | pH 6.8, 65°C, recombinant wild-type enzyme | Aquifex aeolicus | |
Metals/Ions
| Metals/Ions | Comment | Organism | Structure |
|---|---|---|---|
| Mg2+ | - |
Aquifex aeolicus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-leucine + tRNALeu | Aquifex aeolicus | - |
AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Aquifex aeolicus | - |
- |
- |
Purification (Commentary)
| Purification (Comment) | Organism |
|---|---|
| recombinant His-tagged alpha- and beta-subunits from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Aquifex aeolicus |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| ATP + L-leucine + tRNALeu | - |
Aquifex aeolicus | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
| ATP + L-leucine + tRNALeu | recombinant tRNALeu substrate, two peptides of eight and nine amino acid residues in the domain located in the alpha subunit are essential for the enzymes activity | Aquifex aeolicus | AMP + diphosphate + L-leucyl-tRNALeu | - |
? | |
| ATP + L-leucine + tRNALeuGAG | - |
Aquifex aeolicus | AMP + diphosphate + L-leucyl-tRNALeuGAG | - |
? | |
Subunits
| Subunits | Comment | Organism |
|---|---|---|
| dimer | (alphabeta) heterodimer with a large extra domain, the leucine-specific domain, inserted into the catalytic domain, the subunit split site is exactly in the middle of the leucine-specific domain and may have a unique function, two peptides of eight and nine amino acid residues in the domain located in the alpha subunit are essential for the enzymes activity | Aquifex aeolicus |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| Leucyl-tRNA synthetase | - |
Aquifex aeolicus |
| LeuRS | - |
Aquifex aeolicus |
Temperature Optimum [°C]
| Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
|---|---|---|---|
| 37 | 65 | broad temperature optimum | Aquifex aeolicus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.4 | - |
L-leucine | pH 6.8, 65°C, recombinant wild-type enzyme | Aquifex aeolicus | |
| 1.5 | - |
tRNALeuGAG | pH 6.8, 65°C, recombinant wild-type enzyme | Aquifex aeolicus | |
| 1.81 | - |
ATP | pH 6.8, 65°C, recombinant wild-type enzyme | Aquifex aeolicus | |
pH Optimum
| pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
|---|---|---|---|
| 6.8 | - |
assay at | Aquifex aeolicus |
Cofactor
| Cofactor | Comment | Organism | Structure |
|---|---|---|---|
| ATP | - |
Aquifex aeolicus | |
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