Figure 5.
The Rsp5 C2 domain binds to PIs. (A) Equivalent amounts of each GST fusion protein were spotted onto nitrocellulose as a control to demonstrate equivalent levels of each protein in the incubation buffer (bottom). The indicated GST fusion proteins (100 ng/ml) were incubated with PIP strips for 2 h at 4°C. Bound proteins were detected by immunoblotting with anti-GST antibodies. (B) Equivalent amounts (50 ng) of each GST fusion protein (input, top) were incubated with phosphatidylcholine liposomes containing 5% (wt/wt) of the indicated PI. Liposomes were collected by centrifugation, and bound proteins were resolved by SDS-PAGE and detected by immunoblotting with anti-GST antibodies (bound, bottom panel).