Open AccessPrimary structure of murine major histocompatibility complex alloantigens: isolation, biochemical characterization, and preliminary alignment of CNBr fragments from the H-2Ib glycoprotein.
Author(s) -
Bruce M. Ewenstein,
Tosiki Nisizawa,
Hiroshi Uehara,
Stanley G. Nathenson,
John E. Coligan,
Thomas J. Kindt
Publication year - 1978
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.75.6.2909
Subject(s) - papain , cleavage (geology) , glycoprotein , chemistry , peptide sequence , major histocompatibility complex , cyanogen bromide , biochemistry , amino acid , microbiology and biotechnology , protein primary structure , biology , stereochemistry , gene , enzyme , paleontology , fracture (geology)
Radiochemical methodology has been applied in studies directed toward the determination of the amino acid sequence of the murine H-2Kb gene product. Five major CNBr fragments, which comprise the NH2-terminal 80% of the intact glycoprotein and include the H-2Kb fragment isolated by papain cleavage, were isolated and characterized. By means of amino-terminal sequence analysis, homology to other products of the major histocompatibility complex, identification of partial CNBr cleavage products, and localization of the site of papain cleavage, it was possible to align the fragments obtained from the molecule in the following order: the NH2-terminal fragment, designated IIIn, contains 23 residues and is followed by fragment IIIa, which contains 29 residues. Two peptides Ib (Mr 11,500) and Ia (Mr 12,000), each of which contains a carbohydrate moiety, are followed by fragment Ic (Mr 5800), which includes the site of papain cleavage. The data indicate the presence of disulfide bonds between fragments Ib and Ia and between Ia and Ic.