Figure 4. IncE108-132 interferes with CI-MPR binding to SNX5-associated complexes.
Lysates from HEK293T cells transiently expressing full-length FLAG-SNX5WT were incubated with anti-FLAG beads in the presence of the indicated concentrations of wild type or mutant IncE108-132 under non-equilibrium conditions. Eluates were immunoblotted with the indicated antibodies. Input represents 1% of lysates. WT but not mutant IncE interferes with binding of CI-MPR to FLAG-SNX5-containing complexes. The data shown is representative of three biological experiments.
Figure 4—figure supplement 1. IncE peptide interferes with IncE101-132 binding to SNX5-PX20-180 in vitro.
Purified 6xHis-MBP-IncE101-132-Strep was immobilized to Strep-Tactin beads and incubated with purified 8xHis-SNX5-PX20-180 in the presence of the indicated concentrations of WT IncE108-132 or IncEV114N,F116D (Mut) under non-equilibrium conditions. Eluates were immunoblotted with the indicated antibodies. WT but not mutant IncE peptide interferes with binding of IncE to SNX5-PX.