title: Characterization of a glycoprotein fusogen isolated from Sendai virus.
abstract: After isolation from Sendai virus, the glycoproteins HN and F retained their ability to induce hemagglutination and both heterologous and homologous cell-cell fusion. Both methods for demonstrating cell fusion indicated that the isolated HN and F glycoproteins compared favorably with whole Sendai virus as a fusogen. Conditions affecting the degree of fusion were examined and optimized. Whole virus and isolated glycoprotein preparations were characterized by electron microscopy and by SDS-polyacrylamide gel electrophoresis. Lipid analysis of the glycoprotein preparations by thin layer chromatography and gas chromatography/mass spectrometry indicated that they were partially lipid-depleted during the isolation protocol and the ratio of cholesterol to phospholipid was higher than in the whole virus. A complete fatty acid analysis was performed on lipid extracts from whole virus and from glycoprotein preparations. Detergent was removed from the glycoproteins by dialysis and by incubation with Amberlite XAD-2 resin. The detergent content of the glycoprotein preparations was monitored by gas chromatography and with [3H]Triton X-100. Both methods showed that virtually all (greater than or equal to 99.8%) of the originally added detergent was removed. Electron microscopy of the negatively-stained HN and F preparations showed primarily spherical particles 120 +/- 20 A in diameter (range 80-250 A). Since no organization reminiscent of envelopes could be demonstrated, we conclude that the fusogenic activity of Sendai virus resides in the glycoproteins per se rather than in bilayer integrated lipid-protein complexes.
mesh_terms: D000818:Animals; D002459:Cell Fusion / Q000187:drug effects; D002460:Cell Line; D005227:Fatty Acids / Q000032:analysis; D006023:Glycoproteins; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D008055:Lipids / Q000032:analysis; D051379:Mice; D008854:Microscopy, Electron; D017830:Octoxynol; D010222:Parainfluenza Virus 1, Human / Q000032:analysis*; D011092:Polyethylene Glycols; D014759:Viral Envelope Proteins / Q000032:analysis / Q000494:pharmacology*; D014760:Viral Fusion Proteins
chemical_list: D005227:Fatty Acids; D006023:Glycoproteins; D008055:Lipids; D014759:Viral Envelope Proteins; D014760:Viral Fusion Proteins; D011092:Polyethylene Glycols; D017830:Octoxynol