Primary structure of murine major histocompatibility complex alloantigens: completion of the sequence of the amino-terminal 284 residues of H-2Kb.

Abstract

The primary structure of the COOH-terminal cyanogen bromide (CNBr) cleavage fragment Ic (CN-Ic) of the extracellular portion of the murine histocompatibility antigen H-2Kb has been completed. CN-Ic contains a site of papain cleavage which has been utilized for solubilizing H-2Kb by cleaving off the membrane integrating portion of the molecule. The amino acid sequence of CN-Ic has been determined by using peptides recovered after trypsin digestion of CN-Ic before and after blockage of lysine groups with citraconic anhydride. Overlapping sequences for the tryptic fragments were obtained by amino-terminal sequence analysis. The sequence of fragment CN-Ic, which spans residues 229-284 in H-2Kb, is as follows: Glu-Leu-Val-Glu-Thr-Arg-Pro-Ala-Gly-Asp-Gly-Thr-Phe-Gln-Lys-Trp-Ala-Ser-Val-Val-Pro-Leu-Gly-Lys-Glu-Gln-Tyr-Tyr-Thr-Cys-His-Val-Tyr-Gln-Gln-Gly-Leu-Pro-Gln-Pro-Leu-Thr-Leu-Arg-Trp-Asp-Glu-Pro-Pro-Ser-Thr-Val-Ser-Asn-Met. This amino acid sequence determination completes the primary structure of the amino terminal 284 residues of H-2Kb, that portion of this histocompatibility antigen which is external to the cell membrane and which contains antigenic determinants. It was also possible to identify Val-281 as a papain cleavage site within CN-Ic. The completed structure was analyzed solely by radiochemical methods. The structure obtained for H-2Kb is 71% homologous to the reported structure of HLA-B7, a human homologue.