Carbohydrate moieties of rat MHC class I antigens.
The rat major histocompatibility complex class I antigens RT1.Au and RT1.Eu from the u haplotype and RT1. An from the n haplotype were labeled with 14C-asparagine or with 3H-fucose, mannose, galactose, and N-acetylglucosamine. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis showed complete removal of radioactivity from the sugar-labeled antigen heavy chains by digestion with glycopeptidase F, an enzyme that removes N-linked glycans completely. High performance liquid chromatography analysis of the tryptic digests of the mixed sugar-labeled and asparagine-labeled antigens demonstrated that all the sugar-labeled peptides were coincident with asparagine-labeled peptides. The An antigen showed three glycopeptides, each of which had different amounts of sugar radioactivity. The antigens Au and Eu showed two glycopeptides with different amounts of radioactivity but at identical positions in the two antigens. Antigen Eu had an additional glycopeptide with a lower amount of radioactivity. The positions of the glycopeptides from the Au and Eu antigens were different from those of the An antigen. The peptide profiles of the 14C-asparagine-labeled Au and Eu antigens demonstrated distinct differences between the molecules. The results of this study show that: (a) all the glycans on rat class I antigens are N-linked, as they are on H-2 and HLA class I antigens; (b) there are compositional differences among the glycans in each of the three antigens; (c) the glycosylation pattern of the rat class I antigens is similar to that of the mouse class I antigens, which contain two or three glycans, in contrast to that of the human class I antigens, which contain only one glycan; and (d) the antigens Au and Eu from the same haplotype are more closely related to each other than they are to the An antigen.
D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D005805:Genes, MHC Class I; D006649:Histocompatibility Antigens / Q000302:isolation & purification; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D051381:Rats; D014357:Trypsin
D002241:Carbohydrates; D006649:Histocompatibility Antigens; D010446:Peptide Fragments; D014357:Trypsin