Identification of blood group A and B antigens in human glycophorin.
Glycophorin A (GPA), the major sialoglycoprotein of human erythrocyte membranes, was isolated separately from blood group A and B erythrocytes using phenol-water extraction. After purification, performed as gel filtration in the presence of SDS, two glycophorin samples GPA-A and GPA-B were run, in duplicate, in SDS-PAGE and electroblotted onto Immobilon P. After staining with 1) anti-glycophorin antibody and 2) with relevant anti-blood group (A or B) antibody it was shown that the band pattern of the samples in each duplicate was the same. GPA-A and GPA-B samples were also degraded using Carlson degradation (beta-elimination in mild alkaline/strong reducing conditions) and from reaction products the fractions of O-glycans and N-glycans were isolated; they were used in hemagglutination inhibition test. It was shown that both sugar fractions derived from GPA-A did inhibit agglutination of blood group A erythrocytes by anti-A antibody, whereas oligosaccharide fractions derived from GPA-B inhibited agglutination of blood group B erythrocytes by anti-B antibody. These results, obtained using immunochemical methods, confirm the presence of blood group A and B determinants in the carbohydrate moiety of human glycophorin, derived from the blood group A or B erythrocytes, respectively.
Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Wrocław.
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Podbielska
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Krotkiewski
D000017:ABO Blood-Group System / Q000737:chemistry* / Q000276:immunology*; D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000032:analysis; D000939:Epitopes / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification; D004912:Erythrocytes / Q000737:chemistry / Q000276:immunology; D008401:Gas Chromatography-Mass Spectrometry; D006021:Glycophorins / Q000737:chemistry* / Q000276:immunology* / Q000302:isolation & purification; D006385:Hemagglutination Inhibition Tests; D006801:Humans; D007519:Isoantigens / Q000737:chemistry* / Q000276:immunology*; D008969:Molecular Sequence Data
D000017:ABO Blood-Group System; D002241:Carbohydrates; D000939:Epitopes; D006021:Glycophorins; D007519:Isoantigens