FIGURE 2.

Rsp5 HECT C-lobe mediates interaction with Cps1 MVB sorting sequence. A, lysates from yeast expressing either TAP-tagged wild type or the indicated WW-domain mutant Rsp5 were incubated with the indicated GST constructs to determine if Cps1 is a WW-domain interacting substrate. Bound TAP-Rsp5 was detected with anti-actin polyclonal antibody following gel electrophoresis. B, protein-protein interaction utilizing the indicated His-MBP-Rsp5 truncations and the indicated bait-GST constructs. Bound material was visualized by anti-MBP Western blotting.