Table 1.
Apparent kinetic constants for incorporation of dGTP and GCV-TP by WT and mutant Pols
| dGTP | GCV-TP | |||||
| Pol | Motif | Km, μM | kcat, min−1 | Km, μM | kcat, min−1 | Ki, μM |
| WT | 0.44 ± 0.05 | 22 ± 0.70 | 5.5 ± 1.2 | 1.7 ± 0.17 | 4.5 ± 0.50 | |
| D301N | Exo I | 0.45 ± 0.09 | 11 ± 0.60 | 7.8 ± 1.5 | 1.0 ± 0.07 | 4.4 ± 0.60 |
| F412V | Exo II | 0.45 ± 0.06 | 5.0 ± 0.20 | 5.7 ± 1.8 | 1.0 ± 0.07 | 5.1 ± 0.90 |
| L545S | Exo III | 0.46 ± 0.07 | 20 ± 0.80 | 5.8 ± 1.3 | 0.87 ± 0.06 | 4.8 ± 0.70 |
Apparent Km values were calculated from assays using radiolabeled primer template by fitting data points to the Michaelis–Menten equation using GraphPad Prism (Version 6). Apparent kcat values were determined by dividing apparent Vmax values by the enzyme concentrations. Apparent Ki values were derived from Lineweaver–Burk plots. Data are mean ± SEs on the basis of two independent replicates.