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. Author manuscript; available in PMC: 2017 Oct 17.
Published in final edited form as: Chem Res Toxicol. 2016 Sep 21;29(10):1741–1754. doi: 10.1021/acs.chemrestox.6b00244

Table 2.

Steady-State Kinetic Parameters for dNTP Incorporation Opposite G by Human Wild-Type Pol κ (1–526) and Variants

pol κ(1–526) dNTP Km (µM) kcat (s−1) kcat/Km (s−1 µM−1) finsa relative
efficiencyb
wild-type A 220 ± 50 0.047 ± 0.003 2.2 × 10−4 0.0024
T 510 ± 100 0.17 ± 0.01 3.3 × 10−4 0.0037
G 71 ± 10 0.040 ± 0.002 5.6 × 10−4 0.0062
C 12 ± 1 1.1 ± 0.03 9.0 × 10−2 1 1
E29K A 250 ± 80 0.067 ± 0.007 2.7 × 10−4 0.0037
T 590 ± 60 0.20 ± 0.01 3.4 × 10−4 0.0048
G 150 ± 30 0.063 ± 0.003 4.2 × 10−4 0.0058
C 14 ± 3 1.0 ± 0.1 7.2 × 10−2 1 0.80
T44M A 220 ± 3 0.010 ± 0.001 4.8 × 10−5 0.00062
T 330 ± 70 0.026 ± 0.002 7.8 × 10−5 0.0010
G 55 ± 6 0.0053 ± 0.0001 1.0 × 10−4 0.0013
C 12 ± 1 0.93 ± 0.03 7.7 × 10−2 1 0.86
F192C A 180 ± 30 0.067 ± 0.003 3.7 × 10−4 0.0044
T 470 ± 60 0.34 ± 0.01 7.2 × 10−4 0.0085
G 110 ± 10 0.089 ± 0.003 8.1 × 10−4 0.0096
C 13 ± 3 1.1 ± 0.1 8.4 × 10−2 1 0.93
R246X A 150 ± 2 0.015 ± 0.001 1.0 × 10−4 0.0075
T 490 ± 10 0.052 ± 0.002 1.1 × 10−4 0.0077
G 92 ± 27 0.011 ± 0.001 1.2 × 10−4 0.0088
C 16 ± 3 0.22 ± 0.01 1.4 × 10−2 1 0.15
E292K A 150 ± 30 0.060 ± 0.003 4.0 × 10−4 0.0046
T 540 ± 50 0.22 ± 0.01 4.0 × 10−4 0.0046
G 79 ± 8 0.050 ± 0.001 6.4 × 10−4 0.0073
C 13 ± 1 1.1 ± 0.03 8.7 × 10−2 1 0.97
R298H A 530 ± 170 0.026 ± 0.002 4.9 × 10−5 0.0060
T 1400 ± 500 0.032 ± 0.004 2.3 × 10−5 0.0028
G 710 ± 190 0.018 ± 0.002 2.6 × 10−5 0.0031
C 75 ± 10 0.62 ± 0.02 8.2 × 10−3 1 0.091
A471V A 280 ± 70 0.033 ± 0.003 1.2 × 10−4 0.0034
T 520 ± 120 0.19 ± 0.01 3.6 × 10−4 0.010
G 110 ± 20 0.036 ± 0.002 3.2 × 10−4 0.0091
C 26 ± 5 0.90 ± 0.06 3.5 × 10−2 1 0.39
T473A A 350 ± 90 0.021 ± 0.002 5.9× 10−5 0.0056
T 920 ± 130 0.040 ± 0.002 4.4× 10−5 0.0041
G 230 ± 70 0.021 ± 0.002 9.0× 10−5 0.0085
C 48 ± 13 0.51 ± 0.04 1.1× 10−2 1 0.12
R512W A 550 ± 60 0.0086 ± 0.0003 1.6 × 10−5 0.0027
T 410 ± 90 0.051 ± 0.003 1.3 × 10−4 0.022
G 300 ± 30 0.030 ± 0.001 1.0 × 10−4 0.017
C 80 ± 24 0.46 ± 0.04 5.8× 10−3 1 0.064
a

Misinsertion frequency, calculated by dividing kcat/Kmfor each dNTP incorporation by the kcat/Km for dCTP incorporation opposite G.

b

Relative efficiency, calculated by dividing kcat/Km of each pol κ(1–526) for dCTP incorporation opposite G by kcat/Km of wild type pol κ(1–526) for dCTP incorporation opposite undamaged G.