Crystallization (Comment) | Organism |
---|---|
crystals of a C-terminally truncated (D20) recombinant Dm-dNK mutant E52D are grown using the vapour diffusion method by hanging drop geometry. The E52D mutant is crystallized with its feedback inhibitor dTTP. The backbone conformation remains unchanged, and coordination between D52 and the dTTPMg complex is observed | Drosophila melanogaster |
Protein Variants | Comment | Organism |
---|---|---|
E52D | markedly decreased kcat | Drosophila melanogaster |
E52H | markedly decreased kcat | Drosophila melanogaster |
E52Q | inactive mutant protein | Drosophila melanogaster |
Q81N | mutation shows a 200fold and 100fold increase in Km. kcat is decreased 5fold and 2fold for dThd and dCyd | Drosophila melanogaster |
R105H | markedly decreased kcat | Drosophila melanogaster |
R105K | inactive mutant protein | Drosophila melanogaster |
Y70W | mutant enzyme loses activity towards purines. Negative cooperativity towards dThd and dCyd is observed | Drosophila melanogaster |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0012 | - |
2'-deoxythymidine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.0022 | - |
5-(E)-(2-bromvinyl)-2'-deoxyuridine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.0023 | - |
2'-deoxycytidine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.0037 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme E52D | Drosophila melanogaster | |
0.0037 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme E52H | Drosophila melanogaster | |
0.0038 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme E52D | Drosophila melanogaster | |
0.0049 | - |
5-(E)-(2-bromvinyl)-2'-deoxyuridine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
0.0058 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme E52H | Drosophila melanogaster | |
0.0089 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme R105H | Drosophila melanogaster | |
0.0243 | - |
1-(beta-D-arabinofuranosyl)-cytosine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.062 | - |
1-(beta-D-arabinofuranosyl)-thymine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.113 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme R105H | Drosophila melanogaster | |
0.205 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme Q81N | Drosophila melanogaster | |
0.231 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme Q81N | Drosophila melanogaster | |
0.246 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
0.251 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
0.357 | - |
1-(beta-D-arabinofuranosyl)-thymine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
1.441 | - |
1-(beta-D-arabinofuranosyl)-cytosine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Drosophila melanogaster | - |
- |
- |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
2'-deoxycytidine + ATP | - |
Drosophila melanogaster | 2'-deoxycytidine 5'-phosphate + ADP | - |
? | |
ATP + 1-(beta-D-arabinofuranosyl)-cytosine | - |
Drosophila melanogaster | ADP + 1-(beta-D-arabinofuranosyl)-cytosine 5'-phosphate | - |
? | |
ATP + 1-(beta-D-arabinofuranosyl)-thymine | - |
Drosophila melanogaster | ADP + 1-(beta-D-arabinofuranosyl)-thymine 5'-phosphate | - |
? | |
ATP + 2'-deoxythymidine | - |
Drosophila melanogaster | ADP + 2'-deoxythymidine 5'-phosphate | - |
? | |
ATP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine | - |
Drosophila melanogaster | ADP + 5-(E)-(2-bromvinyl)-2'-deoxyuridine 5'-phosphate | - |
? |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0003 | - |
1-(beta-D-arabinofuranosyl)-thymine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.00071 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme E52D | Drosophila melanogaster | |
0.00072 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme E52D | Drosophila melanogaster | |
0.0012 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme E52H | Drosophila melanogaster | |
0.0029 | - |
1-(beta-D-arabinofuranosyl)-cytosine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.0059 | - |
5-(E)-(2-bromvinyl)-2'-deoxyuridine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.0067 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme R105H | Drosophila melanogaster | |
0.012 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme E52H | Drosophila melanogaster | |
0.0142 | - |
2'-deoxythymidine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.0162 | - |
2'-deoxycytidine | 37°C, pH 7.5, wild-type enzyme | Drosophila melanogaster | |
0.055 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme R105H | Drosophila melanogaster | |
0.4 | - |
5-(E)-(2-bromvinyl)-2'-deoxyuridine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
1 | - |
1-(beta-D-arabinofuranosyl)-thymine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
2.1 | - |
1-(beta-D-arabinofuranosyl)-cytosine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
2.3 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster | |
5.4 | - |
2'-deoxythymidine | 37°C, pH 7.5, mutant enzyme Q81N | Drosophila melanogaster | |
6.6 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme Q81N | Drosophila melanogaster | |
6.8 | - |
2'-deoxycytidine | 37°C, pH 7.5, mutant enzyme Y70W | Drosophila melanogaster |