Glycophorins of human erythroleukemic K562 cells.
Glycophorins related to alpha glycophorin, of the human erythrocyte membrane, were isolated from human erythroleukemic K562 cells. The glycophorins were purified using sodium dodecyl sulfate (SDS)/trichloroacetic acid fractionation and Folch and hot phenol extractions. 0.1-0.2 micrograms was obtained/10(8) cells, or approximately a 15% yield. SDS-gel electrophoresis revealed a pattern similar to erythrocyte alpha glycophorin except for the slower mobility of the glycophorin monomer. Two populations of K562 glycophorins, present in nearly equivalent amounts, were distinguished by their binding to Lens culinaris lectin agarose. The two populations exhibited similar gel electrophoretic patterns except for the presence of delta-like glycophorin exclusively in the population that did not bind to L. culinaris lectin. Immunoblotting revealed a lack of reaction of the major alpha and delta-like glycophorin bands in all K562 glycophorins with M or N erythrocyte glycophorin-specific monoclonal antibodies. Only minor species of intermediate electrophoretic mobility in glycophorins not binding to L. culinaris showed a reaction with these antibodies. Both populations of glycophorins incorporated radiolabeled glucosamine, mannose, and fucose and contained O-glycosidically linked tri- and tetrasaccharides, present in a ratio of approximately 1:1 indicating a significant degree of hyposialylation when compared to erythrocyte alpha glycophorin. No precursor/product relationship was demonstrated between the major forms of two populations. K562 cell surface labeling with lactoperoxidase revealed that only the glycophorins that exhibited binding to L. culinaris were accessible to iodination and could be the only species expressed at the cell surface.
D001665:Binding Sites; D002460:Cell Line; D002846:Chromatography, Affinity; D004910:Erythrocyte Membrane / Q000032:analysis*; D006021:Glycophorins / Q000302:isolation & purification*; D006412:Hematopoietic Stem Cells / Q000032:analysis*; D006801:Humans; D007120:Immunochemistry; D037102:Lectins; D008715:Methionine / Q000378:metabolism; D009844:Oligosaccharides / Q000032:analysis; D037121:Plant Lectins; D012795:Sialoglycoproteins / Q000302:isolation & purification*
D006021:Glycophorins; D037102:Lectins; D009844:Oligosaccharides; D037121:Plant Lectins; D012795:Sialoglycoproteins; C033619:lentil lectin; D008715:Methionine