Combinatorial Polarized TIRF-AFM Study of Membrane Reorganization by Alpha-Synuclein and Human Prion Protein

Abstract

cells. Using surface plasmon resonance and centrifugation assays, we have found that the Smurf1 C2 domain binds to phosphoinositides and phosphatidylserine in an synergistic fashion. Confocal images of Smurf1 C2-GFP demonstrate that the domain localizes to the plasma membrane as well as intracellular vesicles in cells. Site-directed mutagenesis has shown the specific residues in the loop region of the protein involved in its cellular membrane localization. In addition, we have used a rapamycin-inducible phosphoinositide phosphatase system to demonstrate that this domain binds phosphoinositides at the plasma membrane. We conclude that the unique properties of the Smurf1 C2 domain to sense specific lipids in addition to anionic charge enable it to target multiple subcellular locations.