Table 2.
Pre-Steady-State Kinetic Parameters for wt and qm RB69pol with all 16 dNTP/dN combinations.(Included are two dPuTPs vs templating THF)
| kpol (s−1) | Kd,app (μM) | kpol/Kd,app (μM−1 s−1) | ||||
|---|---|---|---|---|---|---|
|
|
||||||
| dNTP/dN | wt* | Qm | wt* | qm | wt* | qm |
|
| ||||||
| dTTP/dA | 270 | 7 ± 0.1 | 42 | 4 ± 0.5 | 6.4 | 1.7 |
| dATP/dA | 0.13 | 360 ± 28 | 810 | 750 ± 100 | 1.6 × 10−4 | 0.48 |
| dGTP/dA | 0.003 | 400 ± 50 | 780 | 2000 ± 430 | 3.8 × 10−6 | 0.2 |
| dCTP/dA | N.A.D | 26 ± 1 | N.A.D | 160 ± 24 | 1.7 × 10−4 | 0.17 |
|
| ||||||
| dGTP/dC | 160 | 11 ± 0.5 | 66 | 11 ± 2 | 2.4 | 1.0 |
| dATP/dC | 0.08 | 27 ± 0.5 | 1800 | 32 ± 4 | 4.4 × 10−5 | 0.85 |
| dTTP/dC | 0.09 | 15 ± 0.1 | 1700 | 31 ± 1 | 5.2 × 10−5 | 0.5 |
| dCTP/dC | N.A.D | 210 ± 20 | N.A.D | 1460 ± 270 | 3.0 × 10−7 | 0.14 |
|
| ||||||
| dCTP/dG | 200 | 7 ± 0.2 | 70 | 9 ± 1 | 2.9 | 0.8 |
| dATP/dG | 0.004 | 370 ± 9 | 910 | 320 ± 24 | 4.4 × 10−4 | 1.1 |
| dGTP/dG | N.A.D | 215 ± 6 | N.A.D | 390 ± 30 | 1.0 × 10−5 | 0.55 |
| dTTP/dG | N.A.D | 17 ± 0.5 | N.A.D | 69 ± 10 | 7.0 × 10−5 | 0.24 |
|
| ||||||
| dATP/dT | 220 | 8 ± 0.2 | 50 | 8 ± 0.9 | 4.4 | 1.0 |
| dCTP/dT | N.A.D | 14 ± 0.3 | N.A.D | 31 ± 3 | 6.0 × 10−6 | 0.45 |
| dTTP/dT | 0.015 | 5 ± 0.05 | 1400 | 16 ± 1 | 1.0 × 10−5 | 0.32 |
| dGTP/dT | 0.04 | 8 ± 0.6 | 730 | 650 ± 90 | 5.4 × 10−5 | 0.01 |
|
| ||||||
| dATP/THF | 0.3 | 14 ± 0.4 | 1400 | 5 ± 0.7 | 2.1 × 10−4 | 2.8 |
| dGTP/THF | 0.17 | 11 ± 1 | 1800 | 8 ± 1 | 9.4 × 10−5 | 1.4 |
|
| ||||||
*
N.A.D stands for not accurately determined, because the corresponding Kd,app value is greater than 2 mM, therefore only the kpol/Kd,app values is reliable. The kpol and Kd,app values for dNMP incorporation opposite dN by wt RB69pol were from Zhang.H.17 THF stands for tetrahydrofuran templating site (abasic site).