Solubilization of the H-2 antigen from hybrid cells of mouse mammary ascites tumor and mouse L cells at high cell concentrations in detergent.
Radioiodinated membrane components of L-FM3ANo.2 cells, which were hybrid cells of C3H/He mouse mammary ascites tumor and mouse L cells, were solubilized with non-ionic detergent Nonidet P-40 and immunoprecipitated with anti-H-2k sera. The immune complexes were reduced and subjected to SDS-polyacrylamide gel electrophoresis. When the cells were lysed at a concentration of 2 X 10(7) cells/ml in 0.5% NP-40 buffer, only H-2 molecules including beta2-microglobulin were precipitated, but when the cells were lysed at a concentration of 8 X 10(7) cells/ml in 0.5% NP-40 buffer, two labeled components corresponding to molecules of 105,000 daltons and 85,000 daltons, respectively, were observed in addition to the characteristic radioactive peaks of the H-2 antigen. Dilution of the latter detergent lysate with phosphate buffered saline exhibited no effect on the detection of the two additional components, while further treatment of the lysate with NP-40 buffer abrogated them to be detected. The results indicate that the H-2 molecules were solubilized as physical complexes with certain membrane components of L-FM3ANo.2 cells when the cells were lysed at high cell concentrations in detergent, and suggest that the H-2 molecules and certain membrane components may be physically associated with each other on the cell surface of L-FM3ANo.2 cells.
D000818:Animals; D000951:Antigens, Neoplasm / Q000032:analysis*; D002462:Cell Membrane / Q000032:analysis; D003902:Detergents / Q000494:pharmacology*; D005260:Female; D006183:H-2 Antigens / Q000032:analysis*; D006822:Hybrid Cells / Q000032:analysis*; D007739:L Cells / Q000032:analysis*; D008325:Mammary Neoplasms, Experimental / Q000276:immunology*; D008565:Membrane Proteins / Q000032:analysis; D051379:Mice; D012995:Solubility
D000951:Antigens, Neoplasm; D003902:Detergents; D006183:H-2 Antigens; D008565:Membrane Proteins