Interaction in vitro; bait produced as a recombinant fusion protein in bacterial system; prey derived from S2 cell extract.

rb interacts preferentially with a version of Arf79F (Arf1) that mimics the GTP-bound state, over one that mimics the GDP-bound state.

rb interacts with GTP-bound forms of both Arf79f and Arf102f.

The N-terminal amphipathic helix of Arf protein was replaced with 10-His residues. His-tagged bait Arf protein was then used to coat the surface of liposomes containing low levels of a lipid with Ni-nitrilotriacetic acid (NTA) head group. Bait-coated liposomes are mixed with cell lysate, then liposomes are purified by centrifugation. This allows for the identification of protein interactions that occur preferentially or exclusively at the surface of a lipid bilayer.