title: Generation of a water-soluble oligomeric ectodomain of the Rous sarcoma virus envelope glycoprotein.
abstract: Sequences encoding the transmembrane domain of the Rous sarcoma virus envelope (Env) glycoprotein were deleted and replaced with sequences that signal addition of a glycosyl phosphatidylinositol (GPI) membrane anchor. Stable NIH 3T3 cell lines expressing either the wild-type transmembrane-anchored Env or the Env chimera with a GPI tail were established. The GPI-anchored envelope glycoprotein is expressed, oligomerized, and transported to the cell surface in a manner identical to that of its wild-type transmembrane-anchored counterpart. The GPI-linked protein is quantitatively removed from the cell surface by treatment with phosphatidylinositol phospholipase C. The phosphatidylinositol phospholipase C-released, water-soluble Env glycoprotein ectodomain retains the wild-type oligomeric structure and provides a useful tool for studying the subgroup-specific binding and fusion activities of a prototypic retroviral Env glycoprotein.
authors.affiliation: Department of Biochemistry, School of Medicine, University of California, San Francisco 94143-0450.
authors.forename: J M
authors.initials: JM
authors.lastname: Gilbert
authors.forename: L D
authors.initials: LD
authors.lastname: Hernandez
authors.forename: T
authors.initials: T
authors.lastname: Chernov-Rogan
authors.forename: J M
authors.initials: JM
authors.lastname: White
mesh_terms: D016475:3T3 Cells; D000818:Animals; D015703:Antigens, CD / Q000737:chemistry; D001358:Avian Sarcoma Viruses / Q000737:chemistry*; D018958:CD55 Antigens; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins / Q000737:chemistry; D051379:Mice; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D010727:Phosphoric Diester Hydrolases / Q000378:metabolism; D011993:Recombinant Fusion Proteins; D012995:Solubility; D014759:Viral Envelope Proteins / Q000737:chemistry*
chemical_list: D015703:Antigens, CD; D018958:CD55 Antigens; D017261:Glycosylphosphatidylinositols; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; D014759:Viral Envelope Proteins; D010727:Phosphoric Diester Hydrolases; D043265:Phosphatidylinositol Diacylglycerol-Lyase