Figure 4.

Concentration- and time-dependence of HDH V activity. (A) Increasing amounts of HDH V were incubated in the standard 10 µl reaction mixture for 30 min at 37°C. The amount of enzyme in autoradiograms (I) and (II) is indicated at the top of each lane while the substrate concentration remained constant in all the experiments (0.05 nM, 1000 c.p.m.). The right panel shows the quantitative data of (I) and (II). (B) Kinetic of unwinding performed using 100 pM HDH V and the same reaction conditions used for the concentration-dependence experiments. The reaction time (min) is indicated at the top of each lane. Lanes C and D of each autoradiogram are control assays without enzyme and heat-denatured substrate, respectively.