title: Analysis of Arf1 GTPase-Dependent Membrane Binding and Remodeling Using the Exomer Secretory Vesicle Cargo Adaptor.
abstract: Protein-protein and protein-membrane interactions play a critical role in shaping biological membranes through direct physical contact with the membrane surface. This is particularly evident in many steps of membrane trafficking, in which proteins deform the membrane and induce fission to form transport carriers. The small GTPase Arf1 and related proteins have the ability to remodel membranes by insertion of an amphipathic helix into the membrane. Arf1 and the exomer cargo adaptor coordinate cargo sorting into subset of secretory vesicle carriers in the model organism Saccharomyces cerevisiae. Here, we detail the assays we used to explore the cooperative action of Arf1 and exomer to bind and remodel membranes. We expect these methods are broadly applicable to other small GTPase/effector systems where investigation of membrane binding and remodeling is of interest.
authors.affiliation: Department of Molecular Biology and Genetics, Weill Institute for Cell and Molecular Biology, Cornell University, 457 Weill Hall, Ithaca, NY, 14853, USA.|Department of Molecular Biology and Howard Hughes Medical Institute, Princeton University, Princeton, NJ, 08544, USA.
authors.forename: Jon E
authors.initials: JE
authors.lastname: Paczkowski
authors.affiliation: Department of Molecular Biology and Genetics, Weill Institute for Cell and Molecular Biology, Cornell University, 457 Weill Hall, Ithaca, NY, 14853, USA. jcf14@cornell.edu.
authors.forename: J Christopher
authors.initials: JC
authors.lastname: Fromme
mesh_terms: D020823:ADP-Ribosylation Factor 1 / Q000737:chemistry / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology*; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism
chemical_list: D029701:Saccharomyces cerevisiae Proteins; D020823:ADP-Ribosylation Factor 1
keywords: Arf1; Cargo adaptor; Coat protein; GTPase; Membrane binding; Membrane fission; Membrane remodeling; Membrane scission; Membrane trafficking