Table 6.
template | dNTP | amplitude
|
burst rate constant
|
steady-state rate constant
|
|||||
---|---|---|---|---|---|---|---|---|---|
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 | 3.0 ± 0.4 | 22 ± 6 | 0.25 ± 0.03 | 10 ± 5 | ||||
O2-Me-dT | dATP | 0.420 ± 0.047 | 190 ± 60 | 8.9 | 3.3 ± 0.3 | 0.027 ± 0.267 | 78 ± 37 | ||
O2-Me-dT | dCTP | 0.180 ± 0.020 | 601 ± 130 | 0.3 | 2.3 ± 0.6 | 0.013 ± 0.003 | 250 ± 160 | ||
O2-Me-dT | dGTP | 0.093 ± 0.007 | 213 ± 56 | 0.4 | 5.5 ± 1.1 | 217 ± 121 | 25 | 0.017 ± 0.006 | 592 ± 426 |
O2-Me-dT | dTTP | 0.380 ± 0.033 | 104 ± 34 | 3.6 | 3.6 ± 0.2 | 0.023 ± 0.003 | 110 ± 40 | ||
O2-POB-dT | dATP | 0.313 ± 0.020 | 90 ± 20 | 3.5 | 3.7 ± 0.2 | 43 ± 12 | 86 | 0.017 ± 0.001 | 80 ± 20 |
O2-POB-dT | dCTP | 0.187 ± 0.013 | 300 ± 53 | 0.6 | 2.53 ± 0.05 | 27 ± 3 | 94 | 0.004 ± 0.001 | 8 ± 15 |
O2-POB-dT | dGTP | 0.147 ± 0.013 | 421 ± 105 | 0.3 | 2.42 ± 0.24 | 45 ± 22 | 54 | 0.007 ± 0.001 | 186 ± 63 |
O2-POB-dT | dTTP | 0.307 ± 0.033 | 82 ± 31 | 3.7 | 3.7 ± 0.3 | 42 ± 18 | 88 | 0.024 ± 0.004 | 170 ± 90 |
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 obtain amplitude (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].