Table 3.

Steady-State Kinetic Parameters for dNTP Incorporation Opposite N²-AnthG 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	110 ± 10	0.0028 ± 0.0001	2.5 × 10−5	0.0012
	T	340 ± 40	0.0087 ± 0.0003	2.6 × 10−5	0.0013
	G	85 ± 11	0.0024 ± 0.0001	2.8 × 10−5	0.0014
	C	53 ± 7	1.0 ± 0.04	2.0 × 10−2	1	1
E29K	A	200 ± 30	0.0013 ± 0.0001	6.7 × 10−6	0.00046
	T	350 ± 50	0.0055 ± 0.0002	1.6 × 10−5	0.0011
	G	150 ± 10	0.0014 ± 0.00003	9.3 × 10−6	0.00064
	C	87 ± 13	1.3 ± 0.3	1.5 × 10−2	1	0.75
T44M	A	330 ± 80	0.0067 ± 0.0005	2.0 × 10−5	0.0071
	T	380 ± 80	0.0072 ± 0.0006	1.9 × 10−5	0.0068
	G	100 ± 20	0.00064 ± 0.00006	6.4 × 10−6	0.0023
	C	200 ± 20	0.55 ± 0.02	2.8 × 10−3	1	0.14
F192C	A	220 ± 30	0.0028 ± 0.0001	1.3 × 10−5	0.00079
	T	540 ± 60	0.010 ± 0.0004	1.9 × 10−5	0.0012
	G	46 ± 3	0.0019 ± 0.00004	4.1 × 10−5	0.0026
	C	58 ± 16	0.95 ± 0.08	1.6 × 10−2	1	0.80
R246X	A	130 ± 20	0.00074 ± 0.00003	5.7 × 10−6	0.0020
	T	670 ± 190	0.0023 ± 0.0002	3.4 × 10−6	0.0012
	G	93 ± 22	0.00052 ± 0.00004	5.6 × 10−6	0.0020
	C	90 ± 32	0.26 ± 0.03	2.9 × 10−3	1	0.15
E292K	A	220 ± 40	0.0022 ± 0.0002	1.0 × 10−5	0.00043
	T	330 ± 80	0.0086 ± 0.0006	2.6 × 10−5	0.0011
	G	76 ± 13	0.0015 ± 0.0001	2.0 × 10−5	0.00087
	C	44 ± 8	1.0 ± 0.1	2.3 × 10−2	1	1.2
R298H	A	630 ± 130	0.00069 ± 0.00005	1.1 × 10−6	0.00079
	T	780 ± 170	0.0011 ± 0.0001	1.4 × 10−6	0.0010
	G	210 ± 30	0.00065 ± 0.00002	3.1 × 10−6	0.0022
	C	400 ± 60	0.55 ± 0.03	1.4 × 10−3	1	0.070
A471V	A	220 ± 40	0.0019 ± 0.0001	8.5 × 10−6	0.0010
	T	650 ± 120	0.0058 ± 0.0004	9.0 × 10−6	0.0011
	G	210 ± 50	0.0016 ± 0.0001	7.8 × 10−6	0.00092
	C	130 ± 20	1.1 ± 0.1	8.5 × 10−3	1	0.43
T473A	A	410 ± 70	0.0014 ± 0.0001	3.5 × 10−6	0.0014
	T	510 ± 50	0.0015 ± 0.0001	3.0 × 10−6	0.0012
	G	200 ± 30	0.00072 ± 0.00003	3.6 × 10−6	0.0014
	C	210 ± 30	0.54 ± 0.03	2.6 × 10−3	1	0.13
R512W	A	290 ± 60	0.00045 ± 0.00003	1.5 × 10−6	0.0014
	T	620 ± 80	0.0017 ± 0.0001	2.8 × 10−6	0.0026
	G	220 ± 40	0.00023 ± 0.00001	1.1 × 10−6	0.0010
	C	390 ± 80	0.42 ± 0.03	1.1 × 10−3	1	0.055

^aMisinsertion frequency, calculated by dividing k_cat/K_mfor each dNTP incorporation by the k_cat/K_m for dCTP incorporation opposite N²-AnthG.

^bRelative efficiency, calculated by dividing k_cat/K_m of each pol κ(1–526) for dCTP incorporation opposite N²-AnthG by k_cat/K_m of wild type pol κ(1–526) for dCTP incorporation opposite N²-AnthG.