title: Fragmentation of the human transplantation antigen heavy chain by limited proteolysis, acid cleavage, and cyanogen bromide treatment.
abstract: Highly purified, papain-solubilized HLA-A, -B, and -C antigens comprising a mixture of a great number of allelic forms from at least three loci have been fragmented by limited proteolysis, acid cleavage, and cyanogen bromide treatment. Limited proteolysis of 125I-labeled HLA-A, -B, and -C antigens with trypsin, chymotrypsin, thermolysin, and pepsin resulted in the production of two large fragments. One fragment was associated with beta 2-microglobulin and contained all of the carbohydrate. The other fragment, which had a molecular weight of about 13,000, is most probably derived from the COOH-terminal part of the heavy chain. Acid cleavage of the HLA antigen heavy chain gave rise to two main fragments with molecular weights of 22,000 and 11,000. Both fragments contained disulfide bonds. Two minor components, representing further cleavage products of the 22,000-dalton fragment, were also observed. Cleavage of the HLA antigen heavy chain at methionyl residues gave rise to one carbohydrate-containing, cysteine-free 14,000-dalton fragment and one 20,000-dalton fragment that contained all cysteines but no carbohydrate. NH2-terminal amino acid sequence analyses demonstrated that the 22,000-dalton acid cleavage fragment and the 14,000-dalton cyanogen bromide fragment were derived from the NH2-terminal part of the HLA antigen heavy chain.
mesh_terms: D000595:Amino Acid Sequence; D002918:Chymotrypsin; D003488:Cyanogen Bromide; D006649:Histocompatibility Antigens; D006801:Humans; D006868:Hydrolysis; D046911:Macromolecular Substances; D010206:Papain; D010446:Peptide Fragments / Q000032:analysis; D013154:Spleen; D013820:Thermolysin; D014357:Trypsin; D001613:beta 2-Microglobulin
chemical_list: D006649:Histocompatibility Antigens; D046911:Macromolecular Substances; D010446:Peptide Fragments; D001613:beta 2-Microglobulin; D002918:Chymotrypsin; D014357:Trypsin; D010206:Papain; D013820:Thermolysin; D003488:Cyanogen Bromide