. 2014 Aug 27;27(10):1837–1852. doi: 10.1021/tx5002755

Table 5. Steady-State Kinetic Parameters for dNTP Incorporation Opposite G, O⁶-MeG, and N²-EtG by Wild-Type and Variant hPols ι(1-445) in the Presence of 0.15 mM Mn²⁺.

template	pol ι(1–445)	dNTP	K_m (μM)	k_cat (s^–1)	k_cat/K_m (s^–1 mM^–1)	f_ins^a	relative efficiency^b
G	wild-type	C	1.5 ± 0.1	0.11 ± 0.002	73	1	1
		T	3.3 ± 0.2	0.094 ± 0.002	28	0.38
	Δ1–25	C	0.52 ± 0.05	0.036 ± 0.0007	72	1	0.99
		T	1.2 ± 0.1	0.055 ± 0.001	46	0.64
	ΔD17	C	1.4 ± 0.003	0.12 ± 0.003	86	1	1.2
		T	3.6 ± 0.2	0.12 ± 0.002	33	0.38
	R96G	C	11 ± 1	0.028 ± 0.001	2.5	1	0.034
		T	11 ± 1	0.0075 ± 0.0003	0.68	0.27
	I261M	C	1.3 ± 0.08	0.068 ± 0.0009	52	1	0.71
		T	3.0 ± 0.2	0.068 ± 0.001	23	0.44
	E276K	C	1.4 ± 0.08	0.051 ± 0.0006	36	1	0.49
		T	3.4 ± 0.1	0.047 ± 0.0005	14	0.39
	Y374N	C	2.6 ± 0.2	0.14 ± 0.003	54	1	0.74
		T	3.8 ± 0.2	0.095 ± 0.001	25	0.46
O⁶-MeG	wild-type	C	3.9 ± 0.1	0.10 ± 0.001	26	1	1
		T	3.2 ± 0.2	0.069 ± 0.001	22	0.85
	Δ1–25	C	1.6 ± 0.09	0.039 ± 0.0005	24	1	0.93
		T	1.4 ± 0.2	0.027 ± 0.001	19	0.79
	ΔD17	C	3.8 ± 0.2	0.18 ± 0.003	47	1	1.8
		T	2.5 ± 0.4	0.087 ± 0.003	35	0.74
	R96G	C	12 ± 3	0.019 ± 0.002	1.6	1	0.062
		T	4.3 ± 0.6	0.0073 ± 0.0003	1.7	1.1
	I261M	C	2.3 ± 0.1	0.075 ± 0.001	33	1	1.3
		T	4.0 ± 0.5	0.036 ± 0.001	9.0	0.27
	E276K	C	2.5 ± 0.1	0.044 ± 0.0006	18	1	0.69
		T	3.2 ± 0.4	0.038 ± 0.001	12	0.67
	Y374N	C	3.7 ± 0.3	0.077 ± 0.001	21	1	0.81
		T	2.0 ± 0.3	0.050 ± 0.002	25	1.2
N²-EtG	wild-type	C	27 ± 2	0.034 ± 0.001	1.3	1	1
		T	80 ± 5	0.045 ± 0.001	0.56	0.44
	Δ1–25	C	7.5 ± 1.1	0.011 ± 0.001	1.5	1	1.2
		T	29 ± 2	0.018 ± 0.0005	0.62	0.42
	ΔD17	C	13 ± 1	0.025 ± 0.001	1.9	1	1.5
		T	93 ± 12	0.044 ± 0.003	0.47	0.24
	R96G	C	3.7 ± 0.1	0.045 ± 0.0004	12	1	9.2
		T	26 ± 9	0.0058 ± 0.0008	0.22	0.018
	I261M	C	16 ± 2	0.023 ± 0.001	1.4	1	1.1
		T	87 ± 11	0.042 ± 0.002	0.48	0.34
	E276K	C	20 ± 5	0.018 ± 0.002	0.90	1	0.69
		T	92 ± 13	0.042 ± 0.003	0.46	0.51
	Y374N	C	33 ± 3	0.039 ± 0.02	1.2	1	0.92
		T	82 ± 4	0.035 ± 0.001	0.43	0.36

Misinsertion frequency, calculated by dividing k_cat/K_m for each dNTP incorporation by the k_cat/K_m for dCTP incorporation opposite template base. All values are presented to two significant digits.

Relative efficiency, calculated by dividing k_cat/K_m of each pol ι(1–445) for dCTP incorporation opposite template base by k_cat/K_m of wild-type pol ι(1–445) for dCTP incorporation opposite template base.

Table 5. Steady-State Kinetic Parameters for dNTP Incorporation Opposite G, O6-MeG, and N2-EtG by Wild-Type and Variant hPols ι(1-445) in the Presence of 0.15 mM Mn2+.

Table 5. Steady-State Kinetic Parameters for dNTP Incorporation Opposite G, O⁶-MeG, and N²-EtG by Wild-Type and Variant hPols ι(1-445) in the Presence of 0.15 mM Mn²⁺.