Table 1. Steady-state kinetic parameters of accurate base insertion by hpol κ opposite dG in the presence of WRN.
| kcat (min−1) | KM,dNTP (μM) | kcat/KM,dNTP (min−1 μM−1) | |
|---|---|---|---|
| dCMP insertion opposite dG | |||
| No WRN | 14.6 ± 0.4 | 1.3 ± 0.2 | 11.2 |
| WRN1-1432 | 41.7 ± 1.3 | 1.2 ± 0.2 | 34.7 |
| WRN1-1092/E84A | 46.2 ± 2.1 | 1.5 ± 0.4 | 30.8 |
| WRN1-949/E84A | 18.6 ± 0.5 | 1.6 ± 0.3 | 11.6 |
| WRN500-1150 | 27.0 ± 1.0 | 1.2 ± 0.2 | 22.5 |
| WRN949-1092 | 23.2 ± 1.1 | 1.1 ± 0.4 | 21.1 |
| WRN1-333 | 16.3 ± 0.5 | 1.2 ± 0.3 | 13.6 |
| dAMP insertion opposite dG | |||
| No WRN | 0.32 ± 0.02 | 24.5 ± 7.8 | 0.013 |
| WRN1-1432 | 0.35 ± 0.04 | 23.7 ± 7.5 | 0.015 |
For the determination of kinetic values, graphs of product formation versus dNTP concentration were plotted using nonlinear regression analysis (one-site hyperbolic fit) in the GraphPad prism program. The standard error of the fit is reported for each value.