Separation of the Moloney leukemia virus-determined cell surface antigen (MCSA) from known virion proteins associated with the cell membrane.
Cell membranes of Moloney lymphoma cells (YAC, of strain A origin) were solubilized by NP40. The antigenicity of the solubilized protein fraction was assayed by inhibition of the corresponding cytotoxic reaction against YAC target cells. The Moloney leukemia virus (MLV)-determined cell surface antigen (MCSA) was detected with mouse antisera, produced by the repeated inoculation of heavily irradiated YAC cells into syngeneic mice. Virion proteins gp71, p30, p15, p12 and p10 were identified with goat or rabbit antisera against purified Rauscher and Friend leukemia virus proteins. MCSA was found to bind to Con-A--Sepharose and was eluted by mannoside together with H-2A AND GP71. In contrast, p30, p12, p10 and part of p15 and p15(E), were not retained on the column and could be separated from MCSA. Passage of the glycoprotein fraction through Sephadex G-200 led to the separation of MCSA activity from gp71 and H-2A. MCSA eluted between the immunoglobulin (IgG) and the bovine serum albumin (BSA) size markers. MCSA could be also separated from the known viral proteins and from H-2 by velocity centrifugation in sucrose gradients. It sedimented with approximately 6.6 S ahead of gp71 (4.4 S) and H-2 (3.2 S). It is suggested that MCSA may be a glycoprotein with an approximate molecular weight of 110,000 and distinct from the known viral proteins gp71, p30, p15(E), p12, p10 and from H-2.
D000818:Animals; D000956:Antigens, Viral / Q000302:isolation & purification*; D002462:Cell Membrane / Q000276:immunology; D002499:Centrifugation, Density Gradient; D002847:Chromatography, Agarose; D002850:Chromatography, Gel; D003601:Cytotoxicity Tests, Immunologic; D000939:Epitopes; D007106:Immune Sera; D008223:Lymphoma / Q000276:immunology*; D051379:Mice; D008979:Moloney murine leukemia virus / Q000276:immunology*; D009374:Neoplasms, Experimental / Q000276:immunology; D014764:Viral Proteins
D000956:Antigens, Viral; D000939:Epitopes; D007106:Immune Sera; D014764:Viral Proteins