Cloned (Comment) | Organism |
---|---|
recombinant His-tagged wild-type and mutants from overexpressing Escherichia coli BL21 | Methanocaldococcus jannaschii |
Protein Variants | Comment | Organism |
---|---|---|
E103A | unaltered thermostability, no remaining prolylation activity, 5% remaining cysteinylation activity compared to the wild-type enzyme | Methanocaldococcus jannaschii |
P100A | unaltered thermostability, loss of 90% cysteinylation activity, unaltered prolylation activity compared to the wild-type enzyme | Methanocaldococcus jannaschii |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
L-cysteine | competitive inhibition of prolylation. A 40fold excess over L-proline concentration reduces the prolylation activity by 80%, no inhibition of mutant P100A | Methanocaldococcus jannaschii | |
L-proline | competitive inhibition of cysteinylation. A 40fold excess over L-cysteine concentration reduces the cysteinylation activity by over 80%, no inhibition of mutant E103A | Methanocaldococcus jannaschii |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.022 | - |
L-cysteine | aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C | Methanocaldococcus jannaschii | |
0.027 | - |
L-proline | aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C | Methanocaldococcus jannaschii | |
0.049 | - |
ATP | aminoacylation reaction with L-proline, recombinant wild-type enzyme, pH 7.0, 70°C | Methanocaldococcus jannaschii | |
0.06 | - |
ATP | aminoacylation reaction with L-cysteine, recombinant wild-type enzyme, pH 7.0, 70°C | Methanocaldococcus jannaschii |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-proline + tRNAPro | Methanocaldococcus jannaschii | - |
AMP + diphosphate + L-prolyl-tRNAPro | - |
? | |
ATP + L-proline + tRNAPro | Methanocaldococcus jannaschii DSM 2661 | - |
AMP + diphosphate + L-prolyl-tRNAPro | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Methanocaldococcus jannaschii | Q58635 | - |
- |
Methanocaldococcus jannaschii DSM 2661 | Q58635 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + L-cysteine + tRNA | in later work it is shown that cysteine is attached to tRNA(Pro) and thus constitutes a misaminoacylation event and not a dual specificity | Methanocaldococcus jannaschii | AMP + diphosphate + L-cysteinyl-tRNA | - |
? | |
ATP + L-cysteine + tRNA | in later work it is shown that cysteine is attached to tRNA(Pro) and thus constitutes a misaminoacylation event and not a dual specificity | Methanocaldococcus jannaschii DSM 2661 | AMP + diphosphate + L-cysteinyl-tRNA | - |
? | |
ATP + L-proline + tRNAPro | - |
Methanocaldococcus jannaschii | AMP + diphosphate + L-prolyl-tRNAPro | - |
? | |
ATP + L-proline + tRNAPro | - |
Methanocaldococcus jannaschii DSM 2661 | AMP + diphosphate + L-prolyl-tRNAPro | - |
? |
Synonyms | Comment | Organism |
---|---|---|
ProCysRS | misleading | Methanocaldococcus jannaschii |
proline/cysteine-tRNA ligase | misleading | Methanocaldococcus jannaschii |
prolyl-cysteinyl-tRNA synthetase | misleading | Methanocaldococcus jannaschii |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
70 | - |
assay at | Methanocaldococcus jannaschii |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.02 | - |
L-cysteine | aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C | Methanocaldococcus jannaschii | |
0.09 | - |
L-proline | aminoacylation reaction, recombinant wild-type enzyme, pH 7.0, 70°C | Methanocaldococcus jannaschii |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Methanocaldococcus jannaschii |