Figure 2.
The synthetic abasic site analogue 3CAPS is processed by the major human AP-endonuclease APE1. (A) 0.5 pmol of substrate DNA containing enzymatically produced natural AP-sites (AP) or the synthetic analogues 3CAPS and THF were processed by increasing amounts of recombinant APE1 for 5 min and analysed by denaturing gel electrophoresis. (B) Quantitative analysis of AP–site processing with increasing amounts of APE1. (C) For the kinetic analysis of AP–site processing, substrate DNA was incubated with 0.25 fmol recombinant APE1 for the time indicated. Apparent kcat values were calculated by the curve fitting in the linear range (dotted lines). Error bars, SEM of three experiments.