Cloned (Comment) | Organism |
---|---|
expression of the wild-type and mutant enzymes in enzyme-deficient Escherichia coli strain KY895 | Drosophila melanogaster |
Crystallization (Comment) | Organism |
---|---|
purified recombinant mutant N64D in complex with dT or dTTP, counter diffusion and vapour diffusion methods, 20 mg/ml enzyme with 10 mM dT, is mixed with cyrstallization solution containing 0.15 M MES, pH 6.5, 0.3 M lithium sulfate, and 27,5% w/v PEG 2000 monomethylether, or 10 mg/ml enzyme with 5 mM dTTP is mixed with 0.1 M MES, pH 6.5, 0.16 M lithium sulfate, and 25% v/v PEG 2000 monomethylether, 2 weeks, 15°C, X-ray diffraction structure determination and analysis at 3.1 A and 2.2 A resolution, respectively | Drosophila melanogaster |
Protein Variants | Comment | Organism |
---|---|---|
additional information | expression of the enzyme in enzyme-deficient Escherichia coli strain KY895 confers resistnce against 3'-azido-2',3'-didehydro-3'-deoxythymidine at more than 0.1 mM | Drosophila melanogaster |
N45D | site-directed mutagenesis, decreased activity with natural substrates | Drosophila melanogaster |
N45D/N64D | decreased activity with natural substrates, decreased feedback inhibition by dTTP compared to the wild-type enzyme | Drosophila melanogaster |
N64D | site-directed mutagenesis, highly decreased activity with natural substrates, decreased feedback inhibition by dTTP compared to the wild-type enzyme | Drosophila melanogaster |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
dTTP | competitive versus ATP, feedback inhibition, binding at the active site, binding structure of mutant N64D, modelling | Drosophila melanogaster |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0009 | - |
2'-deoxycytidine | recombinant mutant N45D | Drosophila melanogaster | |
0.0009 | - |
2'-deoxythymidine | recombinant mutants N45D | Drosophila melanogaster | |
0.0111 | - |
3'-azido-2',3'-didehydro-3'-deoxythymidine | recombinant mutant N64D | Drosophila melanogaster | |
0.0117 | - |
3'-azido-2',3'-didehydro-3'-deoxythymidine | recombinant mutant N45D | Drosophila melanogaster | |
0.0232 | - |
2'-deoxythymidine | recombinant mutant N64D | Drosophila melanogaster | |
0.118 | - |
2'-deoxycytidine | recombinant mutant N64D | Drosophila melanogaster | |
0.119 | - |
2'-deoxyadenosine | recombinant mutant N45D | Drosophila melanogaster | |
0.412 | - |
2'-deoxyguanosine | recombinant mutant N45D | Drosophila melanogaster | |
3.82 | - |
2'-deoxyadenosine | recombinant mutant N64D | Drosophila melanogaster | |
20.35 | - |
2'-deoxyguanosine | recombinant mutant N64D | Drosophila melanogaster |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Mg2+ | - |
Drosophila melanogaster |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2'-deoxyadenosine | Drosophila melanogaster | - |
ADP + 2'-deoxyadenosine 5'-phosphate | - |
? | |
ATP + 2'-deoxycytidine | Drosophila melanogaster | - |
ADP + 2'-deoxycytidine 5'-phosphate | - |
? | |
ATP + 2'-deoxyguanosine | Drosophila melanogaster | - |
ADP + 2'-deoxyguanosine 5'-phosphate | - |
? | |
ATP + 2'-deoxythymidine | Drosophila melanogaster | - |
ADP + 2'-deoxythymidine 5'-phosphate | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Drosophila melanogaster | - |
- |
- |
Specific Activity Minimum [µmol/min/mg] | Specific Activity Maximum [µmol/min/mg] | Comment | Organism |
---|---|---|---|
additional information | - |
- |
Drosophila melanogaster |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
ATP + 2'-deoxyadenosine | - |
Drosophila melanogaster | ADP + 2'-deoxyadenosine 5'-phosphate | - |
? | |
ATP + 2'-deoxycytidine | - |
Drosophila melanogaster | ADP + 2'-deoxycytidine 5'-phosphate | - |
? | |
ATP + 2'-deoxyguanosine | - |
Drosophila melanogaster | ADP + 2'-deoxyguanosine 5'-phosphate | - |
? | |
ATP + 2'-deoxythymidine | - |
Drosophila melanogaster | ADP + 2'-deoxythymidine 5'-phosphate | - |
? | |
ATP + 2'-deoxythymidine | substrate binding structure of mutant N64D | Drosophila melanogaster | ADP + 2'-deoxythymidine 5'-phosphate | - |
? | |
ATP + 3'-azido-2',3'-didehydro-3'-deoxythymidine | - |
Drosophila melanogaster | ADP + 3'-azido-2',3'-didehydro-3'-deoxythymidine 5'-phosphate | - |
? | |
ATP + aciclovir | - |
Drosophila melanogaster | ADP + aciclovir phosphate | - |
? |
Synonyms | Comment | Organism |
---|---|---|
dNK | - |
Drosophila melanogaster |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.03 | - |
3'-azido-2',3'-didehydro-3'-deoxythymidine | recombinant mutant N45D | Drosophila melanogaster | |
0.037 | - |
3'-azido-2',3'-didehydro-3'-deoxythymidine | recombinant mutant N64D | Drosophila melanogaster | |
0.12 | - |
2'-deoxyguanosine | recombinant mutant N64D | Drosophila melanogaster | |
0.4 | - |
2'-deoxyadenosine | recombinant mutant N64D | Drosophila melanogaster | |
0.82 | - |
2'-deoxythymidine | recombinant mutant N64D | Drosophila melanogaster | |
1.3 | - |
2'-deoxyguanosine | recombinant mutant N45D | Drosophila melanogaster | |
1.6 | - |
2'-deoxycytidine | recombinant mutants N45D and N64D | Drosophila melanogaster | |
1.83 | - |
2'-deoxyadenosine | recombinant mutant N45D | Drosophila melanogaster | |
2.6 | - |
2'-deoxythymidine | recombinant mutant N45D | Drosophila melanogaster |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
ATP | - |
Drosophila melanogaster |
Ki Value [mM] | Ki Value maximum [mM] | Inhibitor | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0047 | - |
dTTP | uncompetitive versus 2'-deoxythymidine, recombinant wild-type enzyme | Drosophila melanogaster | |
0.0163 | - |
dTTP | competitive versus ATP, recombinant wild-type enzyme | Drosophila melanogaster | |
0.52 | - |
dTTP | uncompetitive versus 2'-deoxythymidine, recombinant mutant N64D | Drosophila melanogaster | |
0.829 | - |
dTTP | competitive versus ATP, recombinant mutant N64D | Drosophila melanogaster |