Demonstration of structural polymorphism among HLA-DR light chains by two-dimensional gel electrophoresis.
Human HLA-DR antigens were immunoprecipitated from Nonidet P-40 extracts of [35S]methionine-labeled B lymphoblastoid cell lines and compared by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and isoelectric focusing (IEF). Two-dimensional (2-D) gel analyses, combining SDS-PAGE in the first dimension and IEF in the second dimension, revealed that the heavy (alpha) and light (beta) chains of each DRw specificity displays microheterogeneity of charge. However, the pattern of the heavy chain did not vary among different DRw specificities. In contrast, the light chains of different DRw types varied both in apparent size and charge distribution. Removal of sialic acids with neuraminidase or inhibition of glycosylation with tunicamycin reduced the microheterogeneity of both DR subunits. However, the heavy and light chains each still focused as two major bands, suggesting that other post-translational modifications contribute to the microheterogeneity or that there are two nonallelic DR-like molecules. After treatment with either neuraminidase or tunicamycin, the DR light chains, but not the heavy chains, were still structurally polymorphic. The DR light chains of serologically cross-reactive specificities displayed similar 2-D gel patterns suggesting that the structural polymorphism of the DR light chains is the basis for the serologically detected polymorphism of the HLA-DR antigens. Two additional polypeptides were observed in immunoprecipitates of DR antigens. These proteins, designated M1 and M2, both had a basic isoelectric point and were invariant among different cell lines. The protein M1 may be intracellular because it can not be immunoprecipitated from the cell surface.
D000595:Amino Acid Sequence; D002460:Cell Line; D004587:Electrophoresis, Agar Gel; D000939:Epitopes; D006649:Histocompatibility Antigens; D006801:Humans; D007147:Immunoglobulin Light Chains; D008214:Lymphocytes / Q000276:immunology; D009439:Neuraminidase; D009844:Oligosaccharides / Q000032:analysis; D011110:Polymorphism, Genetic; D013211:Staphylococcus aureus / Q000276:immunology
D000939:Epitopes; D006649:Histocompatibility Antigens; D007147:Immunoglobulin Light Chains; D009844:Oligosaccharides; D009439:Neuraminidase