Table 2.
Steady-state kinetics results for exo- RB69 polymerases
| Terminus | dNTP: Template | Enzyme | kcat (s−1) | Km (μM) | kcat/Km (s−1 M−1) | f efficiency | kcat/Km F,G/L | f F,G/L |
|---|---|---|---|---|---|---|---|---|
| G-C | dATP:T | L− | 0.12 ± 0.04 | 3.4 (±0.5) | 3.5 ± 0.7 × 104 | 1 | ||
| F− | 0.19 ± 0.13 | 3.8 (±0.8) | 6.5 ± 3.0 × 104 | 1 | 1.9 | |||
| G− | 0.20 ± 0.07 | 4.2 (±0.7) | 4.6 ± 1.0 × 104 | 1 | 1.3 | |||
| G-C | dTTP:T | L− | 0.0031 ± 0.001 | 1000 (±310) | 3.1 ± 0.5 | 9.0 × 10−5 | ||
| F− | 0.19 ± 0.13 | 1500 (±700) | 1.2 ± 0.3 × 102 | 1.8 × 10−3 | 37 | 20 | ||
| G− | 0.16 ± 0.01 | 910 (±110) | 1.8 ± 0.1 × 102 | 3.8 × 10−3 | 56 | 43 | ||
| T-A | dGTP:C | L− | 0.11 ± 0.01 | 0.65 (±0.08) | 17 ± 4 × 104 | 1 | ||
| F− | 0.24 ± 0.01 | 0.74 (±0.09) | 33 ± 3 × 104 | 1 | 1.9 | |||
| G− | 0.080 ± 0.007 | 1.8 (±1.0) | 5.6 ± 3.5 × 104 | 1 | 0.32 | |||
| T-T | dGTP:C | L− | 0.0038 ± 0.001 | 880 (±230) | 4.4 ± 1 | 2.6 × 10−5 | ||
| F− | 0.27 ± 0.10 | 420 (±90) | 6.6 ± 1 × 102 | 2.0 × 10−3 | 150 | 77 | ||
| G− | 0.10 ± 0.01 | 860 (±210) | 1.2 ± 0.5 × 102 | 3.7 × 10−3 | 27 | 14 |
The analyses were performed as described in Materials and methods section.