FIGURE 2.

Sus1 mutant proteins bind to Sac3 and Sgf11 with different affinities in vitro. Recombinant GST-Sac3-(573–805)-Cdc31 complex or GST-Sgf11 was immobilized on GSH beads and incubated with the indicated recombinant 6xHis-Sus1 wild-type (WT) or mutant proteins (input). Proteins bound to the beads were eluted with GSH and analyzed by SDS-PAGE (12% gel, MES buffer) and Coomassie staining. Asterisks indicate Sac3 and Sgf11 degradation products, as determined by mass spectrometry. Note that GST-Sac3-(573–805) harbors the entire CID, which binds to both Cdc31 and Sus1.