Cloned (Comment) | Organism |
---|---|
gene alaS, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
G674D | site-directed mutagenesis, a point mutation in the C-terminal domain, the mutation produces a monomeric variant with a fivefold reduced aminoacylation activity compared to the wild-type enzyme | Escherichia coli |
additional information | a truncated enzyme comparising the N-terminal 700 amino acids forms a monomeric protein possessing similar aminoacylation activity like the wild-type enzyme, and construction of N461 truncated enzyme. Aminoacylation activity assay after refolding from denatured state reveals that the monomeric mutants are unable to regain their activity, whereas the dimeric full-length alaRS gets back similar activity as the native enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | thermodynamics of wild-type and mutant enzymes | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | required | Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-alanine + tRNAAla | Escherichia coli | - |
AMP + diphosphate + L-alanyl-tRNAAla | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P00957 | - |
- |
Purification (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutant enzmyes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography | Escherichia coli |
Renatured (Comment) | Organism |
---|---|
after refolding from denatured state, the dimeric full-length alaRS gets back similar activity as the native enzyme, while the monomeric, truncated or point mutants do not, secondary structure analysis, unfolding profiles, overview | Escherichia coli |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-alanine + tRNAAla | - |
Escherichia coli | AMP + diphosphate + L-alanyl-tRNAAla | - |
? |
Subunits | Comment | Organism |
---|---|---|
homodimer | 2 * 96000, analytical ultracentrifugation | Escherichia coli |
More | alanyl-tRNA synthetase exists as a dimer in its native form and the C-terminal coiled-coil part plays an important role in the dimerization process. Dimerization is one of the key regulatory factors that is important in the proper folding and stability of Escherichia coli alaRS | Escherichia coli |
Synonyms | Comment | Organism |
---|---|---|
Alanyl-tRNA synthetase | - |
Escherichia coli |
AlaRS | - |
Escherichia coli |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Escherichia coli |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Escherichia coli |
General Information | Comment | Organism |
---|---|---|
evolution | the enzyme belongs to the class IIa aminoacyl-tRNA synthestase family | Escherichia coli |