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Characterization of Glycophorin A and Band 3 from Tn Polyagglutinable Erythrocytes
Author(s) -
Jokinen Mikko
Publication year - 1981
Publication title -
scandinavian journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.904
H-Index - 84
eISSN - 1600-0609
pISSN - 0036-553X
DOI - 10.1111/j.1600-0609.1981.tb01659.x
Subject(s) - glycophorin , galactose oxidase , gel electrophoresis , chemistry , biochemistry , pronase , lectin , band 3 , polyacrylamide gel electrophoresis , microbiology and biotechnology , sepharose , galactose , trypsin , biology , membrane , enzyme , membrane protein
The 2 types of erythrocytes from a person with persistent mixed‐field polyagglutinability (Tn abnormality) were separated from each other by preparative cell electrophoresis. Surface labelling using the galactose oxidase/NaB 3 H 4 technique followed by polyacrylamide gel electrophoresis showed a strong labelling in the glycophorin A region of Tn positive erythrocytes indicating exposed galactosyl N‐acetyl/galactosaminyl residues. Tn positive cell membranes were labelled by the galactose oxidase/NaB 3 H 4 technique and solubilized in non‐ionic detergent. After chromatography on Helix pomatia lectin‐linked Sepharose, glycophorin A was immunoprecipitated from the sugar eluate using specific antiserum. Glycophorin A from Tn negative cells and normal red blood cells did not bind to Helix pomatia lectin but to Lens culinaris lectin‐Sepharose. Glycophorin A and band 3 were isolated by preparative gel electrophoresis from normal cells and the two red cell populations of the Tn individual. Pronase treatment of labelled glycophorin A followed by gel filtration revealed a more efficient proteolysis in molecules isolated from Tn positive cells. Mild alkaline treatment of galactose oxidase/NaB 3 H 4 or periodate/NaB 3 H 4 labelled glycophorin A liberated 3 different oligosaccharides from Tn positive cells. No significant difference was found between the oligosaccharides of band 3 protein from normal and Tn positive cells and the amounts of glycophorin A were identical in both cell types when determined by radioimmunoassay.