Table 2.

Kinetic parameters for nucleotide incorporation into gapped or recessed DNA catalyzed by Pol λ at 37 °C.

dNTP	kp(s−1)	Kd(µM)	kp/Kd(µM−1s−1)	Efficiency ratioa	Fidelityb
21-19/41-mer (1-nucleotide gap)
dGTP	2.7 ± 0.1	1.9 ± 0.2	1.4	-
dCTP	0.00145 ± 0.00005	1.0 ± 0.1	1.5 × 10−3	-	1.0 × 10−3
dATP	0.00047 ± 0.00002	0.9 ± 0.1	5.2 × 10−4	-	3.7 × 10−4
dTTP	0.00135 ± 0.00009	2.9 ± 0.6	4.7 × 10−4	-	3.3 × 10−4
21-19/42-mer (2-nucleotide gap)
dGTP	1.77 ± 0.02	1.51 ± 0.06	1.2	1
dCTP	0.0161 ± 0.0004	0.69 ± 0.08	2.3 × 10−2	16 ↑	2.0 × 10−2
dATP	0.00037 ± 0.00002	1.2 ± 0.2	3.1 × 10−4	2 ↑	2.6 × 10−4
dTTP	0.00070 ± 0.00003	3.8 ± 0.5	1.8 × 10−4	3 ↑	1.6 × 10−4
21-19/43-mer (3-nucleotide gap)
dGTP	1.99 ± 0.06	1.5 ± 0.2	1.3	1
dCTP	0.0079 ± 0.0002	0.79 ± 0.08	1.0 × 10−2	7 ↑	7.5 × 10−3
dATP	0.00044 ± 0.00003	0.6 ± 0.2	7.3 × 10−4	1	5.5 × 10−4
dTTP	0.00095 ± 0.00008	2.8 ± 0.8	3.4 × 10−4	1	2.6 × 10−4
21-19/44-mer (4-nucleotide gap)
dGTP	1.64 ± 0.03	1.6 ± 0.1	1.0	1
dCTP	0.0247 ± 0.0006	0.8 ± 0.1	3.1 × 10−2	21 ↑	2.9 × 10−2
dATP	0.00126 ± 0.00004	0.48 ± 0.07	2.6 × 10−3	5 ↑	2.6 × 10−3
dTTP	0.00129 ± 0.00007	2.8 ± 0.5	4.6 × 10−4	1	4.5 × 10−4
21-19/45-mer (5-nucleotide gap)
dGTP	2.15 ± 0.06	1.6 ± 0.2	1.3	1
dCTP	0.053 ± 0.001	0.9 ± 0.1	5.9 × 10−2	41 ↑	4.2 × 10−2
dATP	0.0031 ± 0.0001	0.54 ± 0.07	5.7 × 10−3	11 ↑	4.3 × 10−3
dTTP	0.0118 ± 0.0006	2.7 ± 0.4	4.4 × 10−3	9 ↑	3.2 × 10−3
21-19/46-mer (6-nucleotide gap)
dGTP	1.53 ± 0.06	0.7 ± 0.1	2.2	2 ↑
dCTP	0.0450 ± 0.0008	0.79 ± 0.07	5.7 × 10−2	39 ↑	2.5 × 10−2
dATP	0.00149 ± 0.00002	0.46 ± 0.03	3.2 × 10−3	6 ↑	1.5 × 10−3
dTTP	0.0115 ± 0.0003	2.0 ± 0.2	5.8 × 10−3	12 ↑	2.6 × 10−3
21-19/47-mer (7-nucleotide gap)
dGTP	1.86 ± 0.04	1.2 ± 0.1	1.6	1
dCTP	0.049 ± 0.001	0.9 ± 0.1	5.4 × 10−2	38 ↑	3.4 × 10−2
dATP	0.00066 ± 0.00001	0.24 ± 0.02	2.8 × 10−3	5 ↑	1.8 × 10−3
dTTP	0.0051 ± 0.0002	1.1 ± 0.2	4.6 × 10−3	10 ↑	3.0 × 10−3
21-19/48-mer (8-nucleotide gap)
dGTP	1.01 ± 0.03	1.6 ± 0.2	0.6	2 ↓
dCTP	0.0082 ± 0.0002	0.47 ± 0.06	1.7 × 10−2	12 ↑	2.7 × 10−2
dATP	0.00023 ± 0.00001	0.8 ± 0.2	2.9 × 10−4	2 ↓	4.6 × 10−4
dTTP	0.00072 ± 0.00002	5.1 ± 0.4	1.4 × 10−4	3 ↓	2.2 × 10−4
21-19/49-mer (9-nucleotide gap)
dGTP	0.83 ± 0.03	3.0 ± 0.3	0.3	5 ↓
dCTP	0.00226 ± 0.00005	0.38 ± 0.05	5.9 × 10−3	4 ↓	2.1 × 10−2
dATP	No incorporation
dTTP	No incorporation
21-19/50-mer (10-nucleotide gap)
dGTP	0.026 ± 0.001	0.7 ± 0.1	3.7 × 10−2	38 ↓
dCTP	0.00164 ± 0.00007	1.1 ± 0.2	1.5 × 10−3	1	3.9 × 10−2
dATP	No incorporation
dTTP	No incorporation
21/41-mer (no gap)
dGTP	0.0353 ± 0.0008	0.82 ± 0.08	4.3 × 10−2	33 ↓
dCTP	0.00044 ± 0.00003	3.0 ± 0.7	1.5 × 10−4	10 ↓	3.4 × 10−3
dATP	No incorporation
dTTP	No incorporation

^aAn upward-pointing arrow (↑) indicates the ratio was calculated as (k_p/K_d)_{≥2-nucleotide gap}/(k_p/K_d)_{1-nucleotide gap}; a downward-pointing arrow (↓) indicates the calculation used a reciprocal of the equation as follows: (k_p/K_d)_{1-nucleotide gap}/(k_p/K_d)_{≥2-nucleotide gap}.

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