Generation of a water-soluble oligomeric ectodomain of the Rous sarcoma virus envelope glycoprotein.
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.
Gilbert
J M
JM
Department of Biochemistry, School of Medicine, University of California, San Francisco 94143-0450.
Hernandez
L D
LD
Chernov-Rogan
T
T
White
J M
JM
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*
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