High frequency antigens of human erythrocyte membrane sialoglycoproteins. I. Ena receptors in the glycosylated domain of the MN sialoglycoprotein.

The specificity of various allo- and autoantibodies, which agglutinate normal erythrocytes, but do not react with En(a-) red cells and normal erythrocytes, treated with trypsin (anti-EnaTS) or ficin (anti-EnaFS), was investigated. Various fragments and modification products of the major (MN) red cell membranes sialoglycoprotein were used in hemagglutination inhibition assays. Six anti-EnaFS sera were found to be directed against the residues approx. 46-56 of the molecule. Five of these require the carbohydrate unit, attached to Thr50, for binding. One anti-EnaTS serum was found to be directed against the residues approx. 36-42. Another antibody with anti-EnaTS specificity was shown to react with the residues 31-39 in some of the MN sialoglycoprotein molecules, namely those not glycosylated at a certain position (probably Thr33). A third anti-EnaTS serum, directed against the sequence domain around Lys30, was also found to react only with a fraction of the molecules, apparently due to the variable attachment of oligosaccharides in that region. The heterogeneity of glycosylation, detected by these two sera, appears to account for the partial tryptic and chymotryptic cleavage in this domain of the MN sialoglycoprotein, which has been described previously. Heterogeneity of the glycosylation at various positions of the molecule could be established by the isolation and analysis of peptides.