Cloned (Comment) | Organism |
---|---|
overexpression in Escherichia coli strain JM103 | Thermococcus kodakarensis |
overexpression of wild-type isozyme AspRS2 and seleno-Met isozyme AspRS2 in Escherichia coli | Thermus thermophilus |
Crystallization (Comment) | Organism |
---|---|
purified wild-type and seleno-Met isozymes AspRS2, vapour phase diffusion from mother liquid: 100 mM CHES buffer, pH 9.5, 200 mM NaCl, 10% w/v PEG 8000, X-ray diffraction structure determination at 2.3 A resolution, structure analysis | Thermus thermophilus |
Protein Variants | Comment | Organism |
---|---|---|
additional information | wild-type discriminating enzyme is engineered to a non-discriminating mutant by site-directed mutagenesis, a L1 loop exchange | Thermococcus kodakarensis |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.00014 | - |
tRNAAsp | recombinant wild-type enzyme, pH 7.2, 37°C | Thermus thermophilus | |
0.0002 | - |
tRNAAsp | recombinant wild-type enzyme, pH 7.2, 37°C | Thermococcus kodakarensis | |
0.0002 | - |
tRNAAsn | recombinant wild-type enzyme, pH 7.2, 37°C | Thermus thermophilus | |
0.0009 | - |
tRNAAsn | recombinant mutant enzyme, pH 7.2, 37°C | Thermococcus kodakarensis | |
0.001 | - |
tRNAAsp | recombinant mutant enzyme, pH 7.2, 37°C | Thermococcus kodakarensis |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + tRNAAsn | Thermococcus kodakarensis | the archaeal AspRS2 enzyme is discriminating, which means that it forms only Asp-tRNAAsp and not Asp-tRNAAsn, the L1 loop exchange mutant is rendered non-dicriminating | AMP + diphosphate + L-aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | Thermus thermophilus | the archaeal AspRS2 enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn | AMP + diphosphate + L-aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsp | Thermus thermophilus | - |
AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? | |
ATP + L-aspartate + tRNAAsp | Thermococcus kodakarensis | - |
AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Thermococcus kodakarensis | - |
- |
- |
Thermus thermophilus | - |
isozyme AspRS2 | - |
Purification (Comment) | Organism |
---|---|
recombinant from overexpressing Escherichia coli strain JM103, to homogeneity | Thermococcus kodakarensis |
recombinant wild-type and seleno-Met isozyme 2 from overexpressing Escherichia coli | Thermus thermophilus |
Reaction | Comment | Organism | Reaction ID |
---|---|---|---|
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp | determination of structural features for discriminating or nondiscriminating aminoacylation activity | Thermus thermophilus | |
ATP + L-aspartate + tRNAAsp = AMP + diphosphate + L-aspartyl-tRNAAsp | determination of structural features for discriminating or nondiscriminating aminoacylation activity | Thermococcus kodakarensis |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Thermus thermophilus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-aspartate + tRNAAsn | - |
Thermus thermophilus | AMP + diphosphate + L-aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | - |
Thermococcus kodakarensis | AMP + diphosphate + L-aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | the archaeal AspRS2 enzyme is discriminating, which means that it forms only Asp-tRNAAsp and not Asp-tRNAAsn, the L1 loop exchange mutant is rendered non-dicriminating | Thermococcus kodakarensis | AMP + diphosphate + L-aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsn | the archaeal AspRS2 enzyme is nondiscriminating, which means that it forms Asp-tRNAAsp and Asp-tRNAAsn | Thermus thermophilus | AMP + diphosphate + L-aspartyl-tRNAAsn | - |
? | |
ATP + L-aspartate + tRNAAsp | - |
Thermus thermophilus | AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? | |
ATP + L-aspartate + tRNAAsp | - |
Thermococcus kodakarensis | AMP + diphosphate + L-aspartyl-tRNAAsp | - |
? |
Synonyms | Comment | Organism |
---|---|---|
Aspartic acid translase | - |
Thermus thermophilus |
Aspartic acid translase | - |
Thermococcus kodakarensis |
Aspartyl ribonucleate synthetase | - |
Thermus thermophilus |
Aspartyl ribonucleate synthetase | - |
Thermococcus kodakarensis |
aspartyl ribonuleic synthetase | - |
Thermus thermophilus |
aspartyl ribonuleic synthetase | - |
Thermococcus kodakarensis |
Aspartyl-transfer ribonucleic acid synthetase | - |
Thermus thermophilus |
Aspartyl-transfer ribonucleic acid synthetase | - |
Thermococcus kodakarensis |
Aspartyl-transfer RNA synthetase | - |
Thermus thermophilus |
Aspartyl-transfer RNA synthetase | - |
Thermococcus kodakarensis |
Aspartyl-tRNA synthetase | - |
Thermus thermophilus |
Aspartyl-tRNA synthetase | - |
Thermococcus kodakarensis |
AspRS | - |
Thermus thermophilus |
AspRS | - |
Thermococcus kodakarensis |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Thermus thermophilus |
37 | - |
assay at | Thermococcus kodakarensis |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0008 | - |
tRNAAsn | recombinant mutant enzyme, pH 7.2, 37°C | Thermococcus kodakarensis | |
0.0056 | - |
tRNAAsn | recombinant wild-type enzyme, pH 7.2, 37°C | Thermus thermophilus | |
0.0083 | - |
tRNAAsp | recombinant mutant enzyme, pH 7.2, 37°C | Thermococcus kodakarensis | |
0.0117 | - |
tRNAAsp | recombinant wild-type enzyme, pH 7.2, 37°C | Thermococcus kodakarensis | |
0.042 | - |
tRNAAsp | recombinant wild-type enzyme, pH 7.2, 37°C | Thermus thermophilus |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.2 | - |
assay at | Thermus thermophilus |
7.2 | - |
assay at | Thermococcus kodakarensis |