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. 2014 Sep 16;42(18):11408–11418. doi: 10.1093/nar/gku834

Figure 3.

Figure 3.

AP site incision assay of APE1L, APE2 and ARP. (A) Schematic representation of APE1L, APE2 and ARP proteins. EEP, endonuclease-exonuclease-phosphatase; SAP, SAF-A/B, Acinus and PIAS; ZF, GRF-type zinc finger motif. (B) AP endonuclease activity on the AP site. Radiolabeled 35-nt double-stranded DNA containing a THF, an AP site analog, at position 18 (F35[AP]) was used as a substrate for AP endonuclease assay. Reactions were done with 5 nM each of MBP-APE1L, -APE2 and -ARP at 37°C for 30 min. As a reaction control, 0.5 unit of hAPE1 was used. AP endonuclease reaction product (17-nt with 3′-OH) is indicated at the right of the panel. NE, no enzyme control. (C) Kinetics analysis of Arabidopsis AP endonucleases. The incision activity of ARP on AP site was measured by reacting purified MBP-ARP (5 nM) with varying concentrations of substrate (0–100 nM) at 37°C for 4 min. Error bars represent standard deviations from three independent experiments.