Table 1.
Apparent kinetic parameters for the nucleotide- dependent primer modification reaction catalyzed by HIV-1 RT
| Nucleotide | kcat, s−1 | Km, mM | kcat/Km, s−1⋅M−1 |
|---|---|---|---|
| ATP | 0.42 × 10−3 | 1.6 | 0.26 |
| AMP-CPP | 0.042 × 10−3 | 7.8 | 0.0063 |
| AMP-PCP | 0.25 × 10−3 | 10 | 0.024 |
| ADP | 0.28 × 10−3 | 2.0 | 0.14 |
| dATP | 0.27 × 10−3 | 1.6 | 0.17 |
| CTP | 1.2 × 10−3 | 3.1 | 0.39 |
| dCTP | 1.1 × 10−3 | 3.9 | 0.29 |
| GTP | 1.5 × 10−3 | 4.3 | 0.35 |
| dGTP | 0.08 × 10−3 | 0.74 | 0.11 |
| ddGTP | 0.3 × 10−3 | 1.1 | 0.27 |
| UTP | 0.46 × 10−3 | 2.0 | 0.23 |
Primer modification was tested by using [32P]ddAMP-terminated L32 primer and WL50 template in the presence of the indicated PPase-treated nucleotides (0.1–12.8 mM), as described in the legend of Fig. 1. Values represent the average of duplicate experiments.