Isolation and biochemical characterization of the H-2Kk and H-2Dk antigens from the RDM-4 lymphoma.
Milligram amounts of the H-2Kk and H-2Dk antigens from the RDM-4 lymphoma have been isolated in a one-step procedure employing affinity chromatography on monoclonal antibodies conjugated to Sepharose. The purified antigens have been characterized biochemically, including amino-terminal amino acid sequence and tryptic peptide analyses of the 45,000 mol. wt chains of the antigens. The amino acid sequence results for the H-2Kk antigen, when compared with previously reported data for this molecule, reveal a discrepancy in the sequence of this 45,000 mol. wt chain. We postulate that this discrepancy reflects a previously undescribed polymorphism for the Kk-gene. We also report the first amino-terminal amino acid sequence for the 45,000 mol. wt chain from the H-2Dk antigen and demonstrate that the sequence of the beta 2-microglobulin obtained from the H-2Kk antigen preparation is identical to that published previously. Finally, the structural integrity of the H-2Kk antigen is assessed using binding of beta 2-microglobulin, incorporation into liposomes and reaction with antibodies as criteria.
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002460:Cell Line; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006183:H-2 Antigens / Q000302:isolation & purification; D064532:Histocompatibility Antigen H-2D; D008223:Lymphoma / Q000032:analysis*; D051379:Mice; D008970:Molecular Weight; D011863:Radioimmunoassay; D013601:T-Lymphocytes
D000911:Antibodies, Monoclonal; D006183:H-2 Antigens; C024755:H-2K(K) antigen; D064532:Histocompatibility Antigen H-2D