Table 1.
5′-FAM-GGTGGTCCATAAAC 3′-CCACCAGGTATTTGXCTCT-5′ | ||||||
---|---|---|---|---|---|---|
Polymerase | X | dNTP | Km, μM | kcat, min−1 | kcat/Km, μM−1 min−1 | fa |
hPol η | dA | dATP | 3.5 ± 1.4 | 1.7 ± 0.2 | 0.49 ± 0.15 | 0.035 |
dTTP | 1.7 ± 0.3 | 24.5 ± 1.2 | 14.1 ± 2.4 | 1 | ||
dCTP | 3.7 ± 0.6 | 1.4 ± 0.07 | 0.39 ± 0.05 | 0.027 | ||
dGTP | 14.5 ± 2.4 | 2.9 ± 0.13 | 0.21 ± 0.03 | 0.015 | ||
N6-dA-peptide | dATP | 8.5 ± 2.5 | 1.2 ± 0.1 | 0.140 ± 0.03 | 0.099 | |
dTTP | 17 ± 3.4 | 24 ± 1.4 | 1.4 ± 0.2 | 1 | ||
dCTP | 3 ± 0.5 | 0.9 ± 0.04 | 0.32 ± 0.04 | 0.231 | ||
dGTP | 20 ± 2 | 1.5 ± 0.02 | 0.074 ± 0.006 | 0.053 | ||
hPol ⍳ | dA | dTTP | 3 ± 1 | 0.9 ± 0.06 | 0.23 ± 0.08 | 1 |
N6-dA-peptide | dTTP | 2 ± 1.2 | 0.5 ± 0.05 | 0.28 ± 0.16 | 1 | |
hPol κ | dA | dTTP | 1.5 ± 0.3 | 3.2 ± 0.23 | 2.03 ± 0.34 | 1 |
N6-dA-peptide | dTTP | 9.5 ± 4.8 | 0.09 ± 0.008 | 0.009 ± 0.004 | 1 |
Misincorporation frequency (f) = (kcat/Km)incorrect/(kcat/Km)correct.