Effect of AP-endonuclease on wild-type and S326C OGG1

Copyright information:

Taken from "Dimerization and opposite base-dependent catalytic impairment of polymorphic S326C OGG1 glycosylase"

Nucleic Acids Research 2006;34(5):1620-1632.

Published online 20 Mar 2006

PMCID:PMC1405821.

© The Author 2006. Published by Oxford University Press. All rights reserved

() Glycosylase activities of wild-type (black columns) and S326C OGG1 (grey columns) (12.5 nM) were measured with an 8-oxoG·C substrate (250 nM) with or without an equimolar amount of APE1, in the presence or absence of 1 mM DTT. Inset, actual data. Lanes 1–4, wild-type OGG1; lanes 5–8, S326C; lanes 2, 4, 6 and 8, plus 1 mM DTT; lanes 3–4 and 7–8 plus 12.5 nM APE1. Inset, actual data. () Binding of OGG1 enzymes to an abasic site substrate after adding APE1 was measured by EMSA. Wild-type and S326C OGG1 (5 nM) were incubated with an AP site-containing duplex substrate (5 nM) in the presence of 1 mM MgCl at 37°C for 1 min prior to adding an equimolar amount of APE1 and incubating for an additional 1 min at 37°C. Lane 1, no enzyme; lanes 2 and 3, wild-type OGG1; lanes 4 and 5, S326C OGG1; lanes 3 and 5, plus APE1. Inset, actual data. () Borohydride trapping of OGG1 enzymes. Lane 1, molecular weight marker. Trapping of wild-type (lanes 2 and 3) and S326C OGG1 (lanes 4 and 5) with an 8-oxoG·C substrate in the presence (lanes 3 and 5) and absence (lanes 2 and 4) of AP-endonuclease as described in Materials and Methods. Inset, actual data.