. Author manuscript; available in PMC: 2017 Mar 21.

Published in final edited form as: Chem Res Toxicol. 2016 Feb 22;29(3):303–316. doi: 10.1021/acs.chemrestox.5b00468

Table 5.

Kinetic Parameters for the dNTP Dependence of the Pol κ Catalyzed Insertion Opposite O²-alkyl-dT^a

template	dNTP	amplitude			burst rate constant			steady-state rate constant
template	dNTP	A^max/[pol]	K_A (μM)	A^max/[pol]/K_A (mM⁻¹)	k_pol (s⁻¹)	K_d ^dNTP (μM)	k_pol/K_d(s⁻¹ mM⁻¹)	k_ss^max (s⁻¹)	K_ss (μM)
dT	dATP	0.69 ± 0.03	1.9 ± 0.4	363	155 ± 7	7.9 ± 0.8	19600	0.31 ± 0.03	8.0 ± 2.0
O²-Me-dT	dATP	0.72 ± 0.04	22 ± 3	33	7.4 ± 0.4	168 ± 20	44	0.023 ± 0.003	73 ± 41
O²-Me-dT	dCTP	0.22 ± 0.02	100 ± 29	2.2	3.5 ± 0.28	32 ± 15	109	0.014 ± 0.002	130 ± 70
O²-Me-dT	dGTP	0.25 ± 0.02	62 ± 17	4	3.9 ± 0.23	41 ± 12	95	0.013 ± 0.001	41 ± 13
O²-Me-dT	dTTP	0.72 ± 0.04	97 ± 12	7.4	2.6 ± 0.4	96 ± 60	27	0.020 ± 0.002
O²-POB-dT	dATP	0.68 ± 0.04	11 ± 2	62	5.8 ± 0.4	78 ± 16	74	0.026 ± 0.002	55 ± 17
O²-POB-dT	dCTP	0.18 ± 0.02	101 ± 29	1.8	3.89 ± 0.26	51 ± 16	76	0.010 ± 0.002	78 ± 45
O²-POB-dT	dGTP	0.20 ± 0.03	149 ± 67	1.3	3.35 ± 0.45	6 ± 15	555	0.011 ± 0.001	37 ± 14
O²-POB-dT	dTTP	0.72 ± 0.04	97 ± 12	7.4	2.6 ± 0.4	96 ± 60	27	0.020 ± 0.002

Reactions were conducted with 250 nM pol, 25 nM DNA, and 0 to 1000 μM dNTP. The individual time courses were fit to eq 2 to obtain amplitude (A), burst rate constants (k_b), and steady-state rate constants (k_ss) as a function of dNTP concentration. These values were fit to the hyperbolic equation to obtain the parameters in the table. The values are the mean ± standard error of three determinations. If the K values are missing, then the parameter was not dependent on the [dNTP].