. 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 3.

Steady-state kinetic parameters for dNTP incorporation opposite N²-AnthG 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	160 ± 30	0.00074 ± 0.00003	0.0046	0.0014
	T	360 ± 70	0.0015 ± 0.0001	0.0042	0.0013
	G	32 ± 3	0.00029 ± 0.000003	0.0091	0.0028
	C	380 ± 30	1.2 ± 0.03	3.2	1	0.11

L21F	A	400 ± 90	0.00020 ± 0.00001	0.00050	0.00055
	T	510 ± 90	0.0013 ± 0.0001	0.0025	0.0027
	G	73 ± 10	0.00023 ± 0.00001	0.0032	0.0035
	C	1100 ± 68	1.0 ± 0.03	0.91	1	0.031

I39T	A	390 ± 90	0.00039 ± 0.00002	0.0010	0.00059
	T	330 ± 10	0.0012 ± 0.00002	0.0036	0.0021
	G	69 ± 10	0.00020 ± 0.00001	0.0029	0.0017
	C	820 ± 230	1.4 ± 0.17	1.7	1	0.059

D189G	A	450 ± 80	0.000067 ± 0.000003	0.00015	0.00015
	T	520 ± 120	0.0010 ± 0.0001	0.0019	0.0019
	G	510 ± 40	0.00067 ± 0.00002	0.0013	0.0013
	C	1000 ± 200	1.0 ± 0.06	1.0	1	0.034

R219I	A	230 ± 40	0.00053 ± 0.00003	0.0023	0.00068
	T	330 ± 30	0.0024 ± 0.0001	0.0073	0.0021
	G	85 ± 10	0.00053 ± 0.00001	0.0062	0.0018
	C	590 ± 180	2.0 ± 0.27	3.4	1	0.12

E419G	A	760 ± 36	0.0025 ± 0.0001	0.0033	0.024
	T	1100 ± 220	0.0015 ± 0.0002	0.0014	0.010
	G	77 ± 14	0.00011 ± 0.00001	0.0014	0.010
	C	1600 ± 290	0.23 ± 0.01	0.14	1	0.0048

S423R	A	120 ± 10	0.00053 ± 0.00002	0.0044	0.00050
	T	230 ± 40	0.0035 ± 0.0001	0.015	0.0017
	G	27 ± 3	0.00018 ± 0.00001	0.0067	0.00076
	C	170 ± 11	1.5± 0.03	8.8	1	0.30

Y432S	A	600 ± 100	0.00057 ± 0.00003	0.00095	0.0059
	T	520 ± 50	0.0011 ± 0.00003	0.0021	0.013
	G	81 ± 9	0.00029 ± 0.00001	0.0036	0.023
	C	320 ± 110	0.050 ± 0.004	0.16	1	0.0055

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

Relative efficiency, calculated by dividing k_cat/K_m of each pol κ¹⁻⁵²⁶ for dCTP incorporation opposite N²-AnthG by k_cat/K_m of wild type pol κ¹⁻⁵²⁶ for dCTP incorporation opposite undamaged G in Table 2.