Apparent kinetic parameters of nucleotide excision by DNA polymerase ɛ and p53
| Enzyme | Normal 17-mer | 3′-FMdCMP 18-mer | ||||
|---|---|---|---|---|---|---|
| Vmax | Km | Vmax/Km | Vmax | Km | Vmax/Km | |
| nM | nM | |||||
| pol ɛ | 169.1 ± 13.4 | 4.7 ± 0.3 | 36.1 ± 5.0 | 5.0 ± 0.9 | 0.4 ± 0.1 | 14.7 ± 5.6 |
| p 53 | 115.0 ± 11.6 | 26.1 ± 3.3 | 4.4 ± 0.7 | 6.6 ± 0.8 | 2.0 ± 0.8 | 3.7 ± 1.5 |
Various concentrations of the normal 17-base primer/template and 3′-FMdCMP-18-base primer/template were incubated with purified either pol ɛ or p53 in excision reactions for 30 min as described underMaterials and Methods, and the excision products were quantitated. Values were calculated using the Michaelis-Menten equation. Each data point represents the mean ± S.D. of three separate experiments.