Fig. 3.

Investigation of the putative GAPDH binding site on AE1C. GST-tagged WT or mutated AE1C proteins (Δ11; AA1, D⁹⁰²E→AA; YA, Y⁹⁰⁴→A; AA2, D⁹⁰⁵E→AA) or GST alone was employed in bead-bound pull-down assays against GAPDH. A: bound proteins were analyzed by Western blotting using α-GAPDH antibody (with GST blotting providing loading control). B: bands were densitometrically quantified and expressed relative to WT (100%) ± SE. The acidic patch centered on Y⁹⁰⁴ is important for binding. Blots are representative of 3 separate assays. *P = 0.001; +P < 0.001 vs. WT.