Crystallization (Comment) | Organism |
---|---|
recombinant mutant E138D in complex with dUTP, hanging drop vapour diffusion method, 0.002 ml of 2 mg/ml protein in solution with 5 mM dUTP and 20 mM MgSO4 is mixed with 0.002 ml reservoir solution containing 27.5% PEG 400, 50 mM MgSO4 and 0.1 M HEPES, pH 7.5, equilibration against 0.5 ml of reservoir solution, room temeprature, crystal X-ray diffraction structure determination and analysis at 2.1 A resolution | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
E138D | site-directed mutagenesis, the mutant enzyme shows a 140fold reduction in kcat and altered dTTP inhibition compared to the wild-type enzyme | Escherichia coli |
R115Q | site-directed mutagenesis, inactive mutant | Escherichia coli |
S111C | site-directed mutagenesis, inactive mutant | Escherichia coli |
S111T | site-directed mutagenesis, the mutant enzyme shows a 30fold reduction in kcat and altered dTTP inhibition compared to the wild-type enzyme, modeling of the active site of the S111T enzyme, overview | Escherichia coli |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
dTTP | inhibits by increasing the kcat several fold, mutants S111T and E138D are less sensitive to dTTP, overview | Escherichia coli |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Escherichia coli |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
dCTP + H2O | Escherichia coli | - |
dUTP + NH3 | dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP | ? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | P28248 | - |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dCTP + H2O | - |
Escherichia coli | dUTP + NH3 | - |
? | |
dCTP + H2O | - |
Escherichia coli | dUTP + NH3 | dUMP is required for dTMP synthesis via hydrolysis of UTP by dUTPase to dUMP | ? |
Temperature Optimum [°C] | Temperature Optimum Maximum [°C] | Comment | Organism |
---|---|---|---|
37 | - |
assay at | Escherichia coli |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
additional information | - |
additional information | - |
Escherichia coli | |
1.24 | - |
dCTP | pH 6.8, 37°C, recombinant wild-type enzyme | Escherichia coli |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
6.8 | - |
assay at | Escherichia coli |