TABLE 3.
Mutational effects on the rates of single-turnover DNA adenylylation and phosphodiester synthesis
| ChVLig | Step 2 rate constanta | Step 3 rate constantb |
|---|---|---|
| s−1 | s−1 | |
| WT | 2.17 ± 0.16 | 13.6 ± 1.13 |
| R42A | 0.21 ± 0.02 | 1.73 ± 0.21 |
| T43A/K45A | 1.12 ± 0.06 | 8.0 ± 0.45 |
| F75A | 0.035 ± 0.035 | 0.25 ± 0.027 |
| F75L | 0.42 ± 0.025 | 0.39 ± 0.015 |
| M83A | 1.76 ± 0.11 | 7.47 ± 0.41 |
| M83Q | 1.92 ± 0.08 | 18.6 ± 1.69 |
| M83K | 1.24 ± 0.06 | 8.25 ± 0.59 |
| T84A | 2.0 ± 0.09 | 15.4 ± 0.98 |
| F98A | 0.82 ± 0.05 | 2.57 ± 0.24 |
| F98L | 0.50 ± 0.02 | 2.65 ± 0.12 |
| R166K | 2.07 ± 0.08 | 14.0 ± 0.12 |
| K173A | 2.81 ± 0.27 | 10.9 ± 0.74 |
a The step 2 rate constants were derived by fitting the kinetic data in Fig. 4 to a single exponential in Prism.
b The step 3 rate constants were calculated in MATLAB from the kinetic profiles in Figs. 5 and 6 according to the reaction scheme depicted in Fig. 5 (see supplemental Table S1). The rate constants are specified ± the 95% confidence intervals.