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. Author manuscript; available in PMC: 2020 Feb 5.
Published in final edited form as: Chem Res Toxicol. 2014 Apr 21;27(5):919–930. doi: 10.1021/tx500072m

Table 4.

Steady-state kinetic parameters for next base extension from G (or N2-AnthG): C pair template:primer termini by human wild-type and variant pol κ1−526 enzymes

base-pair at 3′ primer termini (template:primer) pol κ1−526 extension with dGTP (the correct nucleotide opposite next template C)
Km (μM) kcat (s−1) kcat/Km (mM−1 s−1) relative extension efficiencya

G:C wild-type 5.0 ± 0.9 0.28 ± 0.02 56 1
L21F 9.0 ± 1.3 0.30 ± 0.02 33 0.59
I39T 6.0 ± 1.3 0.16 ± 0.01 27 0.48
D189G 8.2 ± 0.7 0.40 ± 0.01 49 0.88
R219I 6.0 ± 1.0 0.50 ± 0.03 83 1.5
E419G 23 ± 7 0.30 ± 0.03 13 0.23
S423R 1.4 ± 0.2 0.27 ± 0.01 190 3.4
Y432S 25 ± 4 0.080 ± 0.004 3.2 0.057

N2-AnthG:C wild-type 14 ± 2 0.54 ± 0.02 39 0.70
L21F 65 ± 18 0.41 ± 0.06 6.3 0.11
I39T 97 ± 12 0.60 ± 0.03 6.2 0.11
D189G 200 ± 10 0.60 ± 0.02 3.0 0.054
R219I 7.3 ± 3.0 0.20 ± 0.02 27 0.48
E419G 27 ± 6 0.048 ± 0.004 1.8 0.032
S423R 3.7 ± 0.4 0.12 ± 0.03 32 0.57
Y432S 24 ± 3 0.051 ± 0.003 2.1 0.038
a

Relative extension efficiency, calculated by dividing kcat/Km of each pol κ1−526 for each dGTP incorporation opposite the next template C from the G (or N2-AnthG):C pair by kcat/Km of wild type pol κ1−526 for dGTP incorporation opposite the next template C from the undamaged G:C pair.