TABLE 1.
Steady-state kinetic parameters for nucleotide incorporation by hPolκ with 2,4-difluorotoluene-containing DNA substrates
| DNA substrate | dNTP | kcat (min−1) | Km (μM) | kcat/Km (μM−1 min−1) | Relative efficiency |
|---|---|---|---|---|---|
| 5′-AGA | dTTP | 0.19 ± 0.008 | 0.16 ± 0.03 | 1.2 | 1 |
| 3′-TCTA | dFTP | NDa | ND | ≤1 × 10−5 | ≤8 × 10−6 |
| dATP | 0.036 ± 0.002 | 8.6 ± 3 | 4.2 × 10−3 | 4 × 10−3 | |
| 5′-AGA | dATP | 0.20 ± 0.006 | 0.12 ± 0.02 | 1.7 | 1 |
| 3′-TCTT | dTTP | 0.086 ± 0.004 | 250 ± 30 | 3.4 × 10−4 | 2 × 10−4 |
| 5′-AGA | dATP | 0.095 ± 0.005 | 21 ± 4 | 4.5 × 10−3 | 3 × 10−3 |
| 3′-TCTF | dTTP | 0.11 ± 0.01 | 630 ± 140 | 1.7 × 10−4 | 1 × 10−4 |
a
ND, no nucleotide incorporation detected. Nucleotide incorporation efficiencies are based upon the estimated minimal detectable levels of incorporation.