title: Biophysical and functional assays for viral membrane fusion peptides.
abstract: Membrane fusion is a protein catalyzed biophysical reaction that involves the simultaneous intermixing of two phospholipid bilayers and of the aqueous compartments bound by their respective bilayers. In the case of enveloped virus fusogens, short hydrophobic or amphipathic fusion peptides that are components of the larger fusion complex are essential for the membrane merger event. The process of cell-cell membrane fusion and syncytium formation induced by the nonenveloped fusogenic orthoreoviruses is driven by the Fusion-Associated Small Transmembrane (FAST) proteins, which are similarly dependent on the action of fusion peptides. In this article, we describe some simple methods for the biophysical characterization of viral membrane fusion peptides. Liposomes serve as an ideal model system for characterizing peptide-membrane interactions because their size, shape and composition can be readily manipulated. We present details of fluorescence assays used to elucidate the kinetics of membrane fusion as well as complimentary assays used to characterize peptide-induced liposome binding and aggregation.
authors.affiliation: Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia, Canada B3H4R2.
authors.forename: Jolene A
authors.initials: JA
authors.lastname: Read
authors.forename: Roy
authors.initials: R
authors.lastname: Duncan
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D002467:Cell Nucleus; D004655:Emulsions / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D005470:Fluorometry / Q000295:instrumentation / Q000379:methods; D015726:Giant Cells; D008081:Liposomes / Q000737:chemistry; D050576:Membrane Fusion Proteins; D010743:Phospholipids / Q000737:chemistry; D011485:Protein Binding; D012235:Rhodamines / Q000737:chemistry; D013010:Sonication / Q000379:methods; D013053:Spectrophotometry / Q000379:methods; D014760:Viral Fusion Proteins / Q000737:chemistry*; D053586:Virus Internalization
chemical_list: D004655:Emulsions; D008081:Liposomes; D050576:Membrane Fusion Proteins; D010743:Phospholipids; D012235:Rhodamines; D014760:Viral Fusion Proteins