Table 3.
Kinetic parameters for the interaction of dNTP with P. furiosus polymerase/primer-template complexes.
| This studya | Published work (8)b | |||||
|---|---|---|---|---|---|---|
| Primer-template | kpolc (s-1) |
KDc (μM) |
kpol/KDc (M-1 s-1) |
kpolc (s-1) |
KDc (μM) |
kpol/KDc (M-1 s-1) |
| Thymidine | 2.1 | 9.7 | 1.7 × 105 | 7.7 | 45 | 1.5 × 105 |
| diFBz | 0.2 | 10.5 | 2.0 × 104 | not determined | ||
| FBzIm | 1.6 | 10 | 1.6 × 105 | not determined | ||
| Uracil | too slow for kpol/KD determination | 1.8 × 10-3 | 52 | 35 | ||
| Hypoxanthine | too slow for kpol/KD determination | 2.5 × 10-3 | 51 | 50 | ||
a
Experiments carried out at 37 °C with insertion of a single dGTP
b
Experiments carried out at 50 °C with insertion of a single dATP.
c
The kinetic parameters for the incorporation of a single dNTP into primer-templates (sequences given in the materials and methods) containing the base indicated at +4 were measured under single turnover conditions (figure 6). Values are the averages of three determinations and are accurate to ± 25%.