Table 1.
Steady-state kinetic analysis of insertion opposite N2-dG-15-mer and 36-mer peptide cross-links
| 5′-FAM-GGTGGTCCATAAAC-3′ 3′-CCACCAGGTATTTGXCTCT-5′ | ||||||
|---|---|---|---|---|---|---|
| Polymerase | X | dNTP | kcat, min−1 | Km, μM | kcat/Km, μM−1 min−1 | fa |
| hpol η | dG | dCTP | 9.6 ± 0.4 | 2.7 ± 0.5 | 3.4 ± 0.5 | 1 |
| N2-dG-15-mer peptide cross-link | dCTP | 9.0 ± 0.3 | 0.79 ± 0.15 | 11.4 ± 1.9 | 1 | |
| N2-dG-36-mer peptide cross-link | dCTP | 8.6 ± 0.1 | 0.57 ± 0.05 | 15.0 ± 1.3 | 1 | |
| dG | dATP | 1.3 ± 0.1 | 3.0 ± 1.1 | 0.43 ± 0.13 | 0.13 | |
| N2-dG-15-mer peptide cross-link | dATP | 0.81 ± 0.04 | 2.3 ± 0.8 | 0.34 ± 0.10 | 0.030 | |
| dG | dGTP | 0.94 ± 0.11 | 92 ± 27 | 0.010 ± 0.001 | 0.003 | |
| N2-dG-15-mer peptide cross-link | dGTP | 0.24 ± 0.01 | 4.7 ± 1.1 | 0.05 ± 0.01 | 0.004 | |
| dG | dTTP | 4.5 ± 0.5 | 67 ± 16 | 0.067 ± 0.009 | 0.020 | |
| N2-dG-15-mer peptide cross-link | dTTP | 2.9 ± 0.3 | 78 ± 15 | 0.036 ± 0.04 | 0.003 | |
| hpol κ | dG | dCTP | 2.7 ± 0.3 | 18.7 ± 5.6 | 0.14 ± 0.03 | 1 |
| N2-dG-15-mer peptide cross-link | dCTP | 3.3 ± 0.2 | 3.9 ± 0.8 | 0.84 ± 0.14 | 1 | |
| N2-dG-36-mer peptide cross-link | dCTP | 0.86 ± 0.05 | 5.4 ± 1.3 | 0.15 ± 0.03 | 1 | |
a
Misincorporation frequency (f) = (kcat/Km)incorrect/(kcat/Km)correct.