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. 2008 Nov 7;283(45):30632–30641. doi: 10.1074/jbc.M801401200

FIGURE 3.

FIGURE 3.

GAPDH, but not the variant C152G or C156G, stimulates APE1 AP endonuclease activity. A, increasing concentrations of either purified GST (lanes 9–12) or GST-GAPDH (lanes 13–16) were incubated with purified His-APE1 (lanes 9–16) for 5 min at room temperature before assaying for AP endonuclease activity. Shown are the controls, substrate (80 ng) alone (lane 1), and the following purified proteins Endo IV (0.15 pmol, lane 2), His-APE1 (lanes 3 and 4), GST (lanes 5 and 6), and GST-GAPDH (lanes 7 and 8). The arrows indicate positions of the 42-mer substrate and the 20-mer product. B, increasing amounts (0, 0.16, 0.64, and 0.8 pmol) of either GST-GAPDH (lanes 2–5), or the variants C152G (lanes 6–9), C156G (lanes 10–13), and C247G (lanes 14–17) were incubated with a fixed amount of purified His-APE1 (0.4 pmol each, lanes 1–17) assayed for AP endonuclease activity. Also shown are substrate alone (lane 18) and Endo IV (0.06 pmol, lane 19).