. 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 2.

Steady-state kinetic parameters for dNTP incorporation opposite G by human wild-type and variant pol κ¹⁻⁵²⁶ enzymes

pol κ¹⁻⁵²⁶	dNTP	K_m (μM)	k_cat (s⁻¹)	k_cat/K_m (s⁻¹M⁻¹)	f_ins^a	relative efficiency^b

wild-type	A	470 ± 70	0.0074 ± 0.0003	0.016	0.00055
	T	1600 ± 100	0.19 ± 0.01	0.12	0.0041
	G	150 ± 9	0.029 ± 0.0004	0.19	0.0066
	C	21 ± 3	0.61 ± 0.026	29	1	1

L21F	A	520 ± 90	0.0025 ± 0.0002	0.0048	0.00024
	T	630 ± 160	0.023 ± 0.002	0.037	0.0019
	G	200 ± 40	0.013 ± 0.001	0.065	0.0033
	C	55 ± 9.6	1.1 ± 0.06	20	1	0.69

I39T	A	820 ± 60	0.0078 ± 0.0002	0.0095	0.00043
	T	980 ± 400	0.059 ± 0.01	0.060	0.0027
	G	250 ± 40	0.014 ± 0.001	0.056	0.0025
	C	27 ± 4	1.1 ± 0.06	22	1	0.76

D189G	A	1000 ± 300	0.0067 ± 0.001	0.0067	0.00048
	T	1900 ± 310	0.033 ± 0.003	0.017	0.0012
	G	740 ± 110	0.010 ± 0.001	0.014	0.0010
	C	81 ± 11	1.1 ± 0.05	14	1	0.48

R219I	A	530 ± 80	0.018 ± 0.001	0.034	0.0012
	T	840 ± 60	0.18 ± 0.01	0.21	0.0072
	G	170 ± 30	0.013 ± 0.001	0.076	0.0026
	C	38 ± 3	1.1 ± 0.03	29	1	1.0

E419G	A	900 ± 140	0.0084 ± 0.001	0.0093	0.0072
	T	2200 ± 380	0.017 ± 0.002	0.0077	0.0059
	G	600 ± 110	0.020 ± 0.002	0.033	0.025
	C	150 ± 34	0.19 ± 0.018	1.3	1	0.045

S423R	A	140 ± 30	0.016 ± 0.001	0.11	0.0023
	T	530 ± 60	0.28 ± 0.01	0.53	0.011
	G	93 ± 20	0.016 ± 0.001	0.17	0.0036
	C	17 ± 1.7	0.80 ± 0.02	47	1	1.6

Y432S	A	1500 ± 200	0.014 ± 0.001	0.0093	0.011
	T	1200 ± 300	0.057 ± 0.003	0.0048	0.0056
	G	500 ± 60	0.020 ± 0.001	0.040	0.047
	C	130 ± 19	0.11 ± 0.0066	0.85	1	0.029

Misinsertion frequency, calculated by dividing k_cat/K_m for each dNTP incorporation by the k_cat/K_m for dCTP incorporation opposite G.

Relative efficiency, calculated by dividing k_cat/K_m of each pol κ¹⁻⁵²⁶ for dCTP incorporation opposite G by k_cat/K_m of wild type pol κ^1–526 for dCTP incorporation opposite undamaged G.