TABLE 2.
Kinetic constants for nucleotide incorporation by FMDV WT and M296I 3Dpola
| Template | Nucleotide | WT 3Dpol
|
M296I 3Dpol
|
||||
|---|---|---|---|---|---|---|---|
| Kd,app (μM) | kpol (s−1) | kpol/Kd,app (μM−1 s−1) | Kd,app (μM) | kpol (s−1) | kpol/Kd,app (μM−1 s−1) | ||
| s/s-AU | ATP | 500 | 80 | 0.2 | 400 | 30 | 0.08 |
| GTP | 800 | 0.01 | 1 × 10−5 | 500 | 0.006 | 1 × 10−5 | |
| RTP | 600 | 0.03 | 5 × 10−5 | 1000 | 0.01 | 1 × 10−5 | |
| s/s-AC | GTP | 50 | 200 | 4 | 70 | 100 | 1 |
| RTP | 500 | 0.02 | 4 × 10−5 | 800 | 0.009 | 1 × 10−5 | |
a
All values are rounded to one significant figure. The error in each measurement is no greater than 20%. Experiments were performed as described in Materials and Methods.