Any feedback?
Literature summary for 2.7.1.145 extracted from
- Identification of residues involved in the specificity and regulation of the highly efficient multisubstrate deoxyribonucleoside kinase from Drosophila melanogaster (2000), J. Mol. Biol., 301, 827-837.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| I102M/N117S/M118V/D208N | clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine | Drosophila melanogaster |
| I47T/N210D | clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine | Drosophila melanogaster |
| M1T/T85A/N121S | clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine and 2',3'-dideoxycytidine | Drosophila melanogaster |
| N210D/L239P | clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine | Drosophila melanogaster |
| N38D/N64D | clones harbouring the mutant enzyme are more sensitive specifically to 3'-azido-2',3'-dideoxythymidine | Drosophila melanogaster |
| N45D/N64D | clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and other nucleoside analogs | Drosophila melanogaster |
| N45D/N64D | mutant enzyme with the largest und universal increase in sensitivity of all mutants tested. 316fold increase to 3'-azido-2',3'-dideoxythymidine, more than 11fold to 2',3'-dideoxycytidine, and 3.2fold increase in sensitivity to beta-D-arabinofuranosylcytosine and 2',3'-dideoxyadenosine. Higher Km-values for native substrates than wild-type enzyme and Vmax-values are substantially lower, decrease in feedback inhibition by TTP, Km and Vmax values for 3'-azido-2',3'-dideoxythymidine and Km value for 2',3'-dideoxycytidine are nearly unchanged | Drosophila melanogaster |
| N59D/M118V/Y179H | clones harbouring the mutant enzyme are more sensitive to 3'-azido-2',3'-dideoxythymidine and beta-D-arabinofuranosylcytosine | Drosophila melanogaster |
| N64S/L68S/M69L | clones harbouring the mutant enzyme are more sensitive specifically to 3'-azido-2',3'-dideoxythymidine | Drosophila melanogaster |
| T12A/V84A/N213S | clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine | Drosophila melanogaster |
| T85A | clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine and 2',3'-dideoxycytidine | Drosophila melanogaster |
| V84A | clones harbouring the mutant enzyme are more sensitive to 1-beta-D-arabinofuranosylcytosine | Drosophila melanogaster |
Inhibitors
| Inhibitors | Comment | Organism | Structure |
|---|---|---|---|
| TTP | mutant enzyme N45D/N64D shows a decrease in feedback inhibition compared to the wild-type enzyme | Drosophila melanogaster |  |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.0072 | - |
3'-azido-2',3'-dideoxythymidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 0.0083 | - |
3'-azido-2',3'-dideoxythymidine | - |
Drosophila melanogaster | |
| 0.0242 | - |
thymidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 0.0964 | - |
2'-deoxycytidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 1.093 | - |
2',3'-dideoxycytidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 1.124 | - |
2',3'-dideoxycytidine | - |
Drosophila melanogaster | |
| 2.004 | - |
2'-deoxyguanosine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 3.16 | - |
2'-deoxyadenosine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Drosophila melanogaster | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| (E)-5-(2-bromovinyl)-2'-deoxyuridine + ATP | 54% of the activity with thymidine | Drosophila melanogaster | (E)-5-(2-bromovinyl)-2'-deoxyuridine 5'-phosphate + ADP | - |
? | |
| 1-beta-D-arabinofuranosylcytosine + ATP | 58.2% of the activity with thymidine | Drosophila melanogaster | 1-beta-D-arabinofuranosylcytosine 5'-phosphate + ADP | - |
? | |
| 1-beta-D-arabinofuranosylthymine + ATP | 53.9% of the activity with thymidine | Drosophila melanogaster | 1-beta-D-arabinofuranosylthymine 5'-phosphate + ADP | - |
? | |
| 2',3'-dideoxyadenosine + ATP | 0.9% of the activity with thymidine | Drosophila melanogaster | 2',3'-dideoxyadenosine 5'-phosphate + ADP | - |
? | |
| 2',3'-dideoxycytidine + ATP | - |
Drosophila melanogaster | 2',3'-dideoxycytidine 5'-phosphate + ADP | - |
? | |
| 2'-deoxyadenosine + ATP | - |
Drosophila melanogaster | 2'-deoxyadenosine 5'-phosphate + ADP | - |
? | |
| 2'-deoxycytidine + ATP | - |
Drosophila melanogaster | 2'-deoxycytidine 5'-phosphate + ADP | - |
? | |
| 2'-deoxyguanosine + ATP | - |
Drosophila melanogaster | 2'-deoxyguanosine 5'-phosphate + ADP | - |
? | |
| 2'-deoxyuridine + ATP | 113% of the activity with thymidine | Drosophila melanogaster | 2'-deoxyuridine 5'-phosphate + ADP | - |
? | |
| 2-chloro-2'-deoxyadenosine + ATP | 126% of the activity with thymidine | Drosophila melanogaster | 2-chloro-2'-deoxyadenosine 5'-phosphate + ADP | - |
? | |
| 3'-azido-2',3'-dideoxythymidine + ATP | - |
Drosophila melanogaster | 3'-azido-2',3'-dideoxythymidine 5'-phosphate + ADP | - |
? | |
| 5-fluoro-2'-deoxyuridine + ATP | 92% of the activity with thymidine | Drosophila melanogaster | 5-fluoro-2'-deoxyuridine 5'-phosphate + ADP | - |
? | |
| adenosine + ATP | 0.82% of the activity with thymidine | Drosophila melanogaster | adenosine 5'-phosphate + ADP | - |
? | |
| cytidine + ATP | 16% of the activity with thymidine | Drosophila melanogaster | cytidine 5'-phosphate + ADP | - |
? | |
| guanosine + ATP | 1.18% of the activity with thymidine | Drosophila melanogaster | guanosine 5'-phosphate + ADP | - |
? | |
| thymidine + ATP | - |
Drosophila melanogaster | thymidine 5'-phosphate + ADP | - |
? | |
| uridine + ATP | 6.45% of the activity with thymidine | Drosophila melanogaster | uridine 5'-phosphate + ADP | - |
? | |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.036 | - |
3'-azido-2',3'-dideoxythymidine | - |
Drosophila melanogaster | |
| 0.052 | - |
3'-azido-2',3'-dideoxythymidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 0.076 | - |
2'-deoxyguanosine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 0.54 | - |
2',3'-dideoxycytidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 0.828 | - |
2'-deoxyadenosine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 1.22 | - |
thymidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 4.03 | - |
2'-deoxycytidine | mutant enzyme N45D/N64D | Drosophila melanogaster | |
| 4.2 | - |
2',3'-dideoxycytidine | - |
Drosophila melanogaster | |
| 14.2 | - |
thymidine | pH 7.8, 22°C, recombinant enzyme | Drosophila melanogaster | |
| 15.1 | - |
2'-deoxyguanosine | pH 7.8, 22°C, recombinant enzyme | Drosophila melanogaster | |
| 16.5 | - |
2'-deoxycytidine | pH 7.8, 22°C, recombinant enzyme | Drosophila melanogaster | |
| 20.6 | - |
2'-deoxyadenosine | pH 7.8, 22°C, recombinant enzyme | Drosophila melanogaster | |
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