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. 2010 Aug 3;285(44):34191–34201. doi: 10.1074/jbc.M110.156182

FIGURE 2.

FIGURE 2.

Kinetics of incorporation of dATP and ddATP for the WT and H932Y mutant. For each concentration series, a preformed enzyme-DNA complex ([enzyme] > [DNA duplex]) was rapidly mixed with Mg2+ and various concentrations of dATP or ddATP. In each experiment the final concentrations of the enzyme and DNA after mixing were 150–175 and 75–100 nm, respectively. In globally fitting each data set, the concentration of active enzyme was adjusted to fit the amplitude. A, incorporation of dATP for WT exo− pol γ at various concentrations (0.2 (●), 0.5 (○), 1.5 (■), 3 (□), 5.5 (▴), 8.5 (△) μm) was globally fit to Scheme 1 yielding a kpol of 30 ± 2 s−1 and a Kd,app of 0.7 ± 0.14 μm. B, incorporation of dATP for H932Y exo− pol γ at various concentrations (2.5 (●), 7.5 (○), 20 (■), 40 (□), 100 (▴), 500 (△) μm from bottom to top) was globally fit to Scheme 1 yielding a kpol of 28.6 ± 2.9 s−1 and a Kd,app of 103 ± 15 μm. C, incorporation of ddATP for WT exo− pol γ at various concentrations (0.01 (●), 0.025 (○), 0.05 (■), 0.1 (□), and 5 (▴) μm) was globally fit to Scheme 1 yielding a kpol of 1.8 ± 0.2 s−1 and a Kd,app of 0.42 ± 0.06 μm. D, incorporation of ddATP for H932Y exo− polγ at various concentrations (0.5 (●), 2 (○), 5 (■), 15 (□), and 500 (▴) μm) was globally fit to Scheme 1 yielding a kpol of 1 ± 0.14 s−1 and a Kd,app of 61 ± 10 μm.