Table 7.
template | amplitude
|
burst rate constant
|
steady-state rate constant
|
||||||
---|---|---|---|---|---|---|---|---|---|
dNTP | Amax/[pol] | KA (μM) | Amax/[pol]/KA (mM−1) | kpol (s−1) | Kd dNTP (μM) | kpol/Kd (s−1 mM−1) | kssmax (s−1) | Kss (μM) | |
dT | dATP | 0.3 ± 0.01 | 62 ± 2 | 8 ± 1 | 7750 | 0.86 ± 0.15 | 14 ± 5 | ||
O2-Me-dT | dATP | 0.49 ± 0.11 | 340 ± 190 | 1.4 | 3.3 ± 0.5 | 0.033 ± 0.005 | 202 ± 84 | ||
O2-Me-dT | dCTP | 0.18 ± 0.01 | 320 ± 27 | 0.6 | 3.2 ± 0.7 | 0.013 ± 0.003 | 438 ± 215 | ||
O2-Me-dT | dGTP | 0.14 ± 0.015 | 195 ± 21 | 0.7 | 4.8 ± 0.6 | 210 ± 80 | 22 | 0.010 ± 0.001 | 152 ± 30 |
O2-Me-dT | dTTP | 0.87 ± 0.47 | 1120 ± 960 | 0.7 | 3.2 ± 0.5 | 0.039 ± 0.006 | 364 ± 139 | ||
O2-POB-dT | dATP | 0.29 ± 0.03 | 85 ± 37 | 3.4 | 3.3 ± 0.3 | 20 ± 7 | 165 | 0.029 ± 0.006 | 260 ± 150 |
O2-POB-dT | dCTP | 0.25 ± 0.07 | 860 ± 410 | 0.3 | 2.9 ± 0.4 | 60 ± 40 | 48 | 0.014 ± 0.002 | 490 ± 170 |
O2-POB-dT | dGTP | 0.80 ± 0.67 | 5200 ± 4800 | 0.2 | 2.3 ± 0.1 | 10 ± 4 | 230 | 0.011 ± 0.002 | 380 ± 200 |
O2-POB-dT | dTTP | 0.51 ± 0.03 | 350 ± 60 | 1.5 | 2.8 ± 0.3 | 0.027 ± 0.004 | 110. ± 50 |
Reactions were conducted with 150 nM pol, 15 nM DNA, and 0 to 1000 μM dNTP. The individual time courses were fit to eq 2 to obtainamplitude (A), burst rate constants (kb), and steady-state rate constants (kss) as a function of dNTP concentration. These values were fit to the hyperbolic equation to obtain the parameters in the table. The values are the mean ± standard error of three determinations. If the K values are missing, then the parameter was not dependent on the [dNTP].