Biosynthetic intermediates of HLA class II antigens from B lymphoblastoid cell lines.
Detergent extracts of B lymphoblastoid cell lines (B-LCL) were subjected to immunoaffinity chromatography and gel filtration to purify HLA class II antigens. Class II antigens purified from B-LCL cultured for 24 hr in 10 microM monensin, in which glycoproteins are trapped in transit through the Golgi apparatus, exist in a large macromolecular complex composed of the alpha- and beta-subunits of class II molecules associated with the invariant (I) chain and a sulfated macromolecule that appears to be a proteoglycan. Gel filtration experiments on Sephacryl S-300 reveal that the complex has a Stokes radius corresponding to a globular protein of approximately 270,000 m.w. Analysis of radiolabeled preparations by two-dimensional gel electrophoresis suggests that the complex contains the alpha-, the beta-, and I chain subunits in a 1:1:1 ratio. Dissociation of the protein components followed by gel filtration of the proteoglycan indicates that the proteoglycan contributes approximately 180,000 m.w. to the complex. These results suggest that the complex contains one copy each of the alpha-, the beta-, and the I subunits associated with a proteoglycan molecule. This complex appears to represent a biosynthetic intermediate in the expression of class II molecules which is induced to accumulate intracellularly by monensin treatment of B-LCL.
D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000276:immunology*; D002460:Cell Line; D002846:Chromatography, Affinity; D006680:HLA Antigens / Q000096:biosynthesis* / Q000302:isolation & purification; D006801:Humans; D046911:Macromolecular Substances; D008970:Molecular Weight; D013431:Sulfates / Q000378:metabolism; D013462:Sulfur Radioisotopes
D000911:Antibodies, Monoclonal; D006680:HLA Antigens; D046911:Macromolecular Substances; D013431:Sulfates; D013462:Sulfur Radioisotopes