Cloned (Comment) | Organism |
---|---|
expression of wild-type and mutant TrpRSs in Escherichia coli | Homo sapiens |
Crystallization (Comment) | Organism |
---|---|
purified recombinant enzyme in complex with bovine tRNATrp, hanging drop vapor diffusion method, 4°C, 0.002 ml of protein solution containing 8 mg/ml of TrpRS with an equal amount of tRNA, 10 mM ATP, 1 mM Trp, 20 mM K2HPO4, pH 6.8, 10 mM MgCl2, 5 mM 2-mercaptoethanol and 0.5 mM PMSF, mixed with 0.002 ml reservoir solution containing 2 M (NH4)2SO4 and 50 mM HEPES, pH 7.0, 1-2 months, X-ray diffraction structure determination and analysis at 3.0 A resolution, modeling | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
D99A | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
D99E | site-directed mutagenesis, the mutant shows slightly reduced activity and kcat compared to the wild-type enzyme | Homo sapiens |
D99K | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
D99V | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K102A | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K102D | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K102I | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K102R | site-directed mutagenesis, the mutant shows slightly reduced activity and reduced kcat compared to the wild-type enzyme | Homo sapiens |
K431A | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K431D | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K431I | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
K431R | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
Q194A | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
Q194L | site-directed mutagenesis, the mutant is inactive | Homo sapiens |
R106A | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
R106D | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
R106I | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
R106K | site-directed mutagenesis, the mutant shows slightly reduced activity and reduced kcat compared to the wild-type enzyme | Homo sapiens |
Y159A | site-directed mutagenesis, the mutant shows reduced kcat and activity compared to the wild-type enzyme | Homo sapiens |
Y159F | site-directed mutagenesis, the mutant shows slightly reduced activity and reduce kcat compared to the wild-type enzyme | Homo sapiens |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | kinetics of diverse recombinant mutant enzymes, overview | Homo sapiens | |
0.00121 | - |
tRNATrp | pH 7.5, 30°C, recombinant wild-type enzyme | Homo sapiens |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Homo sapiens |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-tryptophan + tRNATrp | Homo sapiens | - |
AMP + diphosphate + L-tryptophanyl-tRNATrp | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | P23381 | - |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
recombinant wild-type and mutant enzymes | Homo sapiens |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-tryptophan + tRNATrp | - |
Homo sapiens | AMP + diphosphate + L-tryptophanyl-tRNATrp | - |
? | |
ATP + L-tryptophan + tRNATrp | tRNA substrate from Bos taurus, a two step reaction: the amino acid is first activated by ATP to form an aminoacyl-AMP, which is then transferred to the 3' end of the cognate tRNA to form an aminoacyl-tRNA, the discriminator base A73 of the tRNA is specifically recognized by an alpha-helix of the unique N-terminal domain and the anticodon loop by an alpha-helix insertion of the C-terminal domain, the N-terminal domain is involved in Trp activation, but is not essential for tRNA binding and acylation, tryptophan, anticodon, and acceptor arm recognition mechanisms, overview | Homo sapiens | AMP + diphosphate + L-tryptophanyl-tRNATrp | - |
? |
Synonyms | Comment | Organism |
---|---|---|
TrpRS | - |
Homo sapiens |
Tryptophanyl-tRNA synthetase | - |
Homo sapiens |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
30 | - |
assay at | Homo sapiens |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
1.33 | - |
tRNATrp | pH 7.5, 30°C, recombinant wild-type enzyme | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7.5 | - |
assay at | Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Homo sapiens |