Table 3.

Kinetic parameters of nucleotide incorporation into cisplatin-modified DNA

DNA substrate (P/T)	dNTP	kpa(s−1)	Kda (μM)	kp/Kd (μM−1 s−1)	Efficiency ratiob	Fidelityc
21-mer/44DDP	dGTP	6.0 ± 0.2	(2.0 ± 0.2) × 102	3.0 × 10−2	1.5	–
	dATP	0.026 ± 0.002	(1.0 ± 0.1) × 103	2.7 × 10−5	0.97	9.1 × 10−4
	dCTP	0.039 ± 0.007	(6 ± 3) × 102	6.7 × 10−5	2.6	2.3 × 10−3
	dTTP	0.011 ± 0.001	(1.3 ± 0.2) × 103	8.6 × 10−6	1.4	2.9 × 10−4
22-mer/44DDP	dATP	3.8 ± 0.2	(3.9 ± 0.7) × 102	9.8 × 10−3	1.1	–
	dCTP	0.0041 ± 0.0004	(1.4 ± 0.2) × 103	2.9 × 10−6	29	2.9 × 10−4
	dGTP	0.0081 ±.0009	(1.1 ± 0.2) × 103	7.1 × 10−6	10	7.2 × 10−4
	dTTP	0.0021 ± 0.0002	(7 ± 2) × 102	3.2 × 10−6	5.5	3.3 × 10−4
23-mer/44DDP	dCTP	1.6 ± 0.2	(1.3 ± 0.2) × 103	1.3 × 10−3	72	–
	dATP	0.0056 ± 0.0005	(7 ± 1) × 102	8.0 × 10−6	3.4	6.3 × 10−3
	dGTP	0.00050 ± 0.00008	(6 ± 2) × 102	9.1 × 10−7	67	7.2 × 10−4
	dTTP	0.0017 ± 0.0003	(6 ± 2) × 102	2.8 × 10−6	21	2.2 × 10−3
24-mer/44DDP	dCTP	0.043 ± 0.004	(6 ± 1) × 102	7.3 × 10−5	860	–
	dATP	0.00265 ± 0.00009	(4.6 ± 0.4) × 102	5.7 × 10−6	4.7	7.3 × 10−2
	dGTP	0.000171 ± 0.000008	(8.0 ± 0.8) × 102	2.1 × 10−7	290	2.9 × 10−3
	dTTP	0.00075 ± 0.00003	(6.4 ± 0.7) × 102	1.2 × 10−6	51	1.6 × 10−2
25-mer/44DDP	dATP	9.6 ± 0.4	(5.8 ± 0.6) × 102	1.6 × 10−2	6.1	–
	dCTP	0.031 ± 0.004	(1.6 ± 0.3) × 103	1.9 × 10−5	4.4	1.2 × 10−3
	dGTP	0.0115 ± 0.0009	(1.1 ± 0.2) × 103	1.0 × 10−5	6.8	6.3 × 10−4
	dTTP	0.025 ± 0.003	(1.3 ± 0.2) × 103	1.9 × 10−5	0.92	1.2 × 10−3
26-mer/44DDP	dGTP	1.9 ± 0.1	(4.0 ± 0.7) × 102	4.7 × 10−3	36	–
	dATP	0.04 ± 0.01	(1.5 ± 0.8) × 103	2.7 × 10−5	0.98	5.6 × 10−3
	dCTP	0.007 ± 0.002	(1.4 ± 0.5) × 103	5.0 × 10−6	35	1.1 × 10−3
	dTTP	0.0032 ± 0.0006	(1.1 ± 0.4) × 103	2.8 × 10−6	4.3	6.0 × 10−4
27-mer/44DDP	dATP	0.20 ± 0.02	(2.7 ± 0.7) × 102	7.5 × 10−4	17	–
	dCTP	0.0016 ± 0.0003	(8 ± 3) × 102	2.1 × 10−3	40	2.8 × 10−3
28-mer/44DDP	dGTP	4.4 ± 0.3	(5.0 ± 0.9) × 102	8.9 × 10−3	2.8	–
	dATP	0.0057 ± 0.0006	(1.5 ± 0.3) × 103	3.9 × 10−6	6.7	4.3 × 10−4
29-mer/44DDP	dGTP	0.72 ± 0.05	(5.0 ± 0.9) × 102	1.4 × 10−3	4.0	–
	dATP	0.0016 ± 0.0001	(6 ± 1) × 102	2.8 × 10−6	9.4	1.9 × 10−3
30-mer/44DDP	dCTP	3.3 ± 0.2	(2.7 ± 0.4) × 102	1.2 × 10−2	7.1	–
	dGTP	0.0022 ±.0002	(9 ± 2) × 102	2.5 × 10−6	25	2.0 × 10−4
31-mer/44DDP	dTTP	5.5 ± 0.3	(1.5 ± 0.3) × 102	3.7 × 10−2	2.1	–
	dCTP	0.019 ± 0.003	(1.0 ± 0.3) × 103	1.8 × 10−5	0.69	4.9 × 10−4
32-mer/44DDP	dATP	18.9 ± 0.8	(1.3 ± 0.2) × 102	1.5 × 10−1	0.53	–
	dCTP	0.032 ± 0.002	(8 ± 1) × 102	3.9 × 10−5	2.2	2.6 × 10−4

^aThe k_p and K_d obtained with 23-mer/44DDP and 24-mer/44DDP are apparent kinetic parameters due to biphasic kinetics of nucleotide incorporations at these pause sites.

^bCalculated as (k_p/K_d)_control/(k_p/K_d)_damaged in which the (k_p/K_d)_control is derived from Table 4 or ref. (36) for correct and incorrect incorporations, respectively.

^cCalculated as (k_p/K_d)_incorrect/[(k_p/K_d)_correct + (k_p/K_d)_incorrect].