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. 2013 Aug 5;41(19):9077–9089. doi: 10.1093/nar/gkt674

Table 2.

Pre-steady-state kinetic parameters for incorporation of correct and incorrect dNMPs by wt RB69pol and the L415 variants

Pols kpol (s-1) Kd,app (µM) kpol/Kd,app (µM−1 s−1) Discrimination
Correct BP dTTP/dA
    wt 270 42 6.4
    L415A 11 ± 1 2.7 ± 0.6 4.0
    L415G 58 ± 1 9 ± 0.6 6.4
    L415M 375 ± 10 12 ± 1 31.2
Pu-Pu BP dATP/dA
    wt 0.13 810 1.6 × 10−4 4.0 × 104
    L415A 15 ± 1 640 ± 150 2.3 × 10−2 1.7 × 102
    L415G 1 ± 0.01 220 ± 20 4.5 × 10−3 1.4 × 103
    L415M 0.03 ± 0.003 1300 ± 270 2.6 × 10−5 1.2 × 106
Py-Pu BP dCTP/dA
    wt N.A.D >2000 1.7 × 10−4 3.7 × 104
    L415A N.A.D >2000 6.0 × 10−3 6.7 × 102
    L415G 1.6 ± 0.1 1330 ± 200 1.2 × 10 − 3 5.3 × 103
    L415M N.A.D >2000 5.4 × 10−5 5.7 × 105
Py-Py BP dTTP/dC
    wt 0.09 1700 5.2 × 10−5 1.2 × 105
    L415A 130 ± 1 1700 ± 20 7.6 × 10−2 5.3 × 101
    L415G N.A.D >2000 1.1 × 10−2 5.8 × 102
    L415M N.A.D >2000 1.4 × 10−4 2.2 × 105

Pu stands for purine; Py stands for pyrimidine; BP stands for base-pair; N.A.D stands for not accurately determined because the corresponding Kd,app value is greater than 2 mM, and therefore only the kpol/Kd,app values are reliable. Discrimination is defined by (kpol/Kd,app)correct/(kpol/Kd,app)incorrect. Representative gel, progress curve, and plot of kobs versus (dTTP)] are shown in Supplementary Figure S1.