The selective solubilization of different murine splenocyte membrane fractions with lubrol WX and triton X-100 distinguishes two forms of Ia antigens. A cell surface (alpha, beta) and an intracellular (alpha, Ii, beta).
The selective solubilization of different murine lymphocyte membrane compartments with several nonionic detergents was used to study the subcellular distribution of two distinct forms of lymphocyte cell recognition structures (Ia antigens). Ia antigens were isolated with a monoclonal anti-Ia immunoadsorbent from murine splenocytes that had been solubilized with four different nonionic detergents. Analyses of the immunoprecipitates indicated that Lubrol WX was selectively solubilizing a subpopulation of Ia consisting of mature highly glycosylated alpha and beta polypeptides which were not associated with Ii polypeptide. A second Ia subpopulation consisting of less glycosylated cytoplasmic precursor alpha and beta polypeptides associated with Ii polypeptide was immunoprecipitated from the Lubrol WX-insoluble material after solubilizing this material with Triton X-100. Comparable results were obtained when HLA-DR antigens were similarly isolated from cultured human lymphoblastoid cells. This selective solubilization phenomenon was not unique to Ia antigens. Only mature highly glycosylated H-2K molecules were immunoprecipitated from the Lubrol WX-soluble material while the less glycosylated precursor H-2K molecules were immunoprecipitated from the Triton X-100-solubilized Lubrol-insoluble material. These data directly demonstrate that the Ii polypeptide is exclusively associated with the intracellular Ia antigen cytoplasmic precursor molecules. These data also indicate that, under the conditions used in these experiments, Lubrol WX does not completely solubilize integral membrane proteins that have previously been shown to be associated with the rough endoplasmic reticulum.
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000276:immunology; D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II / Q000302:isolation & purification*; D006801:Humans; D007163:Immunosorbent Techniques; D008214:Lymphocytes / Q000276:immunology*; D051379:Mice; D008805:Mice, Inbred A; D008806:Mice, Inbred AKR; D017830:Octoxynol; D011092:Polyethylene Glycols; D012995:Solubility; D013154:Spleen / Q000166:cytology*
D006684:HLA-DR Antigens; D000949:Histocompatibility Antigens Class II; C017474:Lubrol WX; D011092:Polyethylene Glycols; D017830:Octoxynol