Isolation, characterization and amino acid sequence studies of the cyanogen bromide fragments of the H-2Dd glycoprotein.
The papain-solubilized fragment of the H-2Dd antigen, representing the NH2-terminal 80% of the native glycoprotein, can be fragmented into five polypeptides using CNBr cleavage. These fragments have been isolated, characterized and subjected to amino acid sequence analysis using radiochemical microtechniques. It was possible to align these CNBr fragments by comparison of their NH2-terminal amino acid sequence with the known amino acid sequence of the H-2Kb molecule. Thus, the NH2-terminal fragment, D, contains 23 residues and is followed by a glycopeptide, fragment C, of 75 residues. Following fragment C are three fragments held together by disulfide bonds: fragment b4, which has 40 amino acid residues, a second glycopeptide, fragment b2, of 90 residues, and fragment b3, a peptide of approximately 55 residues which terminates at the site of papain cleavage. The complete amino acid sequence of the NH2-terminal CNBr fragment, D, has been determined, and this is presented here along with sequence data for the NH2-terminal 30 residues of each of the other four CNBr fragments of the papain fragment of H-2Dd. A total of 146 positions have been examined in the approximately 280-residue H-2Ddpapain molecule, and a total of 127 residues have been positively determined. Most of the unassigned positions may be tentatively assigned as Asp or Asn, the only amino acids so far not incorporated in radiolabeled form by our metabolic labeling procedures into the H-2Dd molecule. Comparison of the sequence data obtained for the H-2Dd molecule to data available for other histocompatibility antigens revealed homologies of approximately 90% with H-2Kb and H-2Ld, and approximately 75% with HLA-B7 and HLA-A2.
D000595:Amino Acid Sequence; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D002850:Chromatography, Gel; D003488:Cyanogen Bromide; D006023:Glycoproteins / Q000302:isolation & purification*; D006183:H-2 Antigens / Q000302:isolation & purification*; D051379:Mice; D008805:Mice, Inbred A; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D009374:Neoplasms, Experimental / Q000276:immunology; D010206:Papain
D006023:Glycoproteins; D006183:H-2 Antigens; D010206:Papain; D003488:Cyanogen Bromide