TABLE 4.
Steady-state kinetic parameters for next base extension from G (or N2-alkyl G):C (or T) template primer termini by Dpo4
The extent of conversion of primer to the product was kept <20% by adjustment of the enzyme concentration and reaction time.
| Template | Primer | Km, dNTP | kcat × 103 | kcat/Km × 105 | Efficiency relative to G:C |
|---|---|---|---|---|---|
| μm | s-1 | μm-1 s-1 | |||
| G | C | 23 ± 3 | 530 ± 30 | 2300 | |
| T | 180 ± 20 | 17 ± 1 | 10 | 230-fold less | |
| N2-MeG | C | 190 ± 30 | 190 ± 10 | 102 | 23-fold less |
| T | 73 ± 9 | 6.9 ± 0.2 | 9.5 | 240-fold less | |
| N2-EtG | C | 290 ± 60 | 200 ± 20 | 68 | 34-fold less |
| T | 57 ± 9 | 2.6 ± 0.2 | 4.6 | 500-fold less | |
| N2-IbG | C | 820 ± 130 | 260 ± 30 | 31 | 74-fold less |
| T | 110 ± 30 | 0.93 ± 0.05 | 0.8 | 2,900-fold less | |
| N2-BzG | C | 660 ± 140 | 220 ± 30 | 34 | 68-fold less |
| T | 26 ± 4 | 0.70 ± 0.01 | 2.8 | 820-fold less | |
| N2-NaphG | C | 190 ± 30 | 140 ± 10 | 76 | 30-fold less |
| T | 38 ± 9 | 1.4 ± 0.1 | 3.7 | 620-fold less | |
| N2-AnthG | C | 100 ± 20 | 220 ± 20 | 225 | 10-fold less |
| T | 110 ± 20 | 6.7 ± 0.3 | 6.0 | 380-fold less |