title: A quantitative fluorometric approach for measuring the interaction of RhoGDI with membranes and Rho GTPases.
abstract: Tight regulation of Rho GTPase-signaling functions requires the proper localization of proteins to the membrane and cytosolic compartments, which can themselves undergo reconfiguration in response to signaling events. The importance of lipid-mediated membrane signal transduction continues to emerge as a critical event in many Rho GTPase-signaling pathways. Here we describe methods for the reconstitution of lipid-modified Rho GTPases with defined lipid vesicles and how this system can be used as a real-time assay for monitoring protein-membrane interactions.
authors.affiliation: Department of Chemistry and Chemical Biology, Cornell University, Ithaca, NY, USA.
authors.forename: Jared
authors.initials: J
authors.lastname: Johnson
authors.forename: Richard A
authors.initials: RA
authors.lastname: Cerione
authors.forename: Jon W
authors.initials: JW
authors.lastname: Erickson
mesh_terms: D000818:Animals; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005470:Fluorometry / Q000379:methods*; D020730:Guanine Nucleotide Dissociation Inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D008081:Liposomes / Q000737:chemistry / Q000378:metabolism; D008563:Membrane Lipids / Q000235:genetics / Q000378:metabolism*; D011485:Protein Binding; D015398:Signal Transduction; D020741:rho GTP-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors
chemical_list: D020730:Guanine Nucleotide Dissociation Inhibitors; D008081:Liposomes; D008563:Membrane Lipids; D062947:rho-Specific Guanine Nucleotide Dissociation Inhibitors; D020741:rho GTP-Binding Proteins