Fig. 2. Identification of SCAMP2 binding domain within NHE7. GST alone and GST fused to various segments of NHE7 (residues 542-725, 615-725, 666-725, 542-614 and 542-665, as illustrated below) were purified by glutathione-sepharose beads and subjected to GST pull-down. The radiolabeled SCAMP2 protein was incubated with 2 µg purified GST or GST fusion proteins immobilized on glutathione sepharose beads. After extensive washing, bound SCAMP2 was eluted with SDS sample buffer, resolved by SDS-PAGE, and visualized using a PhosphorImager.