Phosphorylation of pyrimidine and deoxycytidine analog monophosphate by human UMP/CMPK
The reactions were performed at 37°C in the presence of 1 mm MgATP and 750 μm phosphate acceptor. Calculations for relative Vmax, Km, and relative efficiency are as mentioned in Table 1. Activity for dCMP phosphorylation was 33 ± 4 μmol/mg/min. Values are presented as mean ± SD from three independent experiments.
| Phosphate acceptor . | Relative Vmax (%) . | Km (μm) . | Relative efficiency (%) . |
|---|---|---|---|
| dCMP | 100 | 500 ± 34 | 100 |
| AraCMP | 827 ± 19 | 917 ± 83 | 451 |
| Gemcitabine monophosphate | 94 ± 10 | 581 ± 25 | 81 |
| d-ddCMP | 10 ± 2 | 272 ± 53 | 18 |
| l-ddCMP | 19 ± 3 | 697 ± 72 | 13 |
| l-(−)-OddCMPa | 19 ± 3 | 1037 ± 52 | 9 |
| l-(−)-SddCMP | 92 ± 10 | 494 ± 94 | 93 |
| l-(−)-FSddCMP | 9 ± 2 | 250 ± 62 | 18 |
| l-(−)-Fd4CMP | 9 ± 2 | 228 ± 68 | 19 |
| Phosphate acceptor . | Relative Vmax (%) . | Km (μm) . | Relative efficiency (%) . |
|---|---|---|---|
| dCMP | 100 | 500 ± 34 | 100 |
| AraCMP | 827 ± 19 | 917 ± 83 | 451 |
| Gemcitabine monophosphate | 94 ± 10 | 581 ± 25 | 81 |
| d-ddCMP | 10 ± 2 | 272 ± 53 | 18 |
| l-ddCMP | 19 ± 3 | 697 ± 72 | 13 |
| l-(−)-OddCMPa | 19 ± 3 | 1037 ± 52 | 9 |
| l-(−)-SddCMP | 92 ± 10 | 494 ± 94 | 93 |
| l-(−)-FSddCMP | 9 ± 2 | 250 ± 62 | 18 |
| l-(−)-Fd4CMP | 9 ± 2 | 228 ± 68 | 19 |
l-(−)-OddCMP, β-l-dioxolane-CMP; l-(−)-Fd4CMP, β-l-2′,3′-dideoxy-2′,3′-didehydro-5-fluoro-CMP.