Cloned (Comment) | Organism |
---|---|
expression of wild-type enzyme and G146A variant in Escherichia coli BL21(DE3)/pDIA17 | Escherichia coli |
Protein Variants | Comment | Organism |
---|---|---|
G146A | mutation is accompanied by a small but significant enhancement of the thermodynamic stability, midpoint denaturation temperature is 3°C higher than that of the wild-type enzyme, midpoint transition is 3.3 M urea, compared to 3.0 M for the wild-type enzyme | Escherichia coli |
General Stability | Organism |
---|---|
midpoint transition is 3.0 M urea for the wild-type enzyme and 3.3 M for the G146A mutant enzyme | Escherichia coli |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.015 | - |
TMP | pH 7.4, 30°C, wild-type enzyme | Escherichia coli | |
0.04 | - |
ATP | pH 7.4, 30°C, independent of cosubstrate | Escherichia coli | |
0.08 | - |
5-bromo-2'-deoxyuridine 5'-monophosphate | pH 7.4, 30°C, wild-type enzyme | Escherichia coli | |
0.25 | - |
dTMP | pH 7.4, 30°C, mutant enzyme G146A | Escherichia coli | |
0.28 | - |
5-bromo-2'-deoxyuridine 5'-monophosphate | pH 7.4, 30°C, mutant enzyme G146A | Escherichia coli | |
2.5 | - |
dUMP | pH 7.4, 30°C, wild-type enzyme | Escherichia coli | |
4 | - |
dUMP | pH 7.4, 30°C, mutant enzyme G146A | Escherichia coli |
Molecular Weight [Da] | Molecular Weight Maximum [Da] | Comment | Organism |
---|---|---|---|
23779 | - |
2 * 23779, electrospray ionization mass spectrometry, wild-type enzyme | Escherichia coli |
23794 | - |
2 * 23794, electrospray ionization mass spectrometry, G146A mutant enzyme | Escherichia coli |
48000 | - |
gel filtration | Escherichia coli |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Escherichia coli | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 5-bromo-2'-deoxyuridine 5'-monophosphate | - |
Escherichia coli | ADP + 5-bromo-2'-deoxyuridine 5'-diphosphate | - |
? | |
ATP + dTMP | - |
Escherichia coli | ADP + dTDP | - |
? | |
ATP + dUMP | - |
Escherichia coli | ADP + dUDP | - |
? |
Subunits | Comment | Organism |
---|---|---|
dimer | 2 * 23779, electrospray ionization mass spectrometry, wild-type enzyme | Escherichia coli |
dimer | 2 * 23794, electrospray ionization mass spectrometry, G146A mutant enzyme | Escherichia coli |
Temperature Stability Minimum [°C] | Temperature Stability Maximum [°C] | Comment | Organism |
---|---|---|---|
57 | - |
midpoint denaturation temperature for wild-type enzyme | Escherichia coli |
60 | - |
midpoint denaturation temperature for the G146A mutant enzyme | Escherichia coli |