Detection of glycophorin A-like glycoproteins on the surface of cultured human cells.
1. In previous studies we have isolated and characterized mucin-type glycopeptides from mouse and human melanoma cells. 2. These glycopeptides have clusters of oligosaccharides of the type (NeuNAc)0-2----[Gal----GalNAc] linked to serine and or threonine suggesting an apparent similarity to glycophorin. 3. We now report the interaction of polyclonal anti-glycophorin antibodies with various cultured cells. Antisera to highly purified glycophorin A were raised in rabbits. 4. Human melanoma cells (HM7), human breast cells (HBL-100) and two lines of human breast cancer cells (MCF-7 and MDA-MB-231) showed medium to very strong cell surface fluorescence pattern after staining with rabbit anti-glycophorin F(ab')2 and FITC-conjugated goat anti-rabbit F(ab')2. 5. Immunodiffusion, immunoelectrophoresis and affinity chromatography on anti-glycophorin IgG-Sepharose 4B of detergent extracts of metabolically labeled cultured cells gave further evidence for the presence of glycophorin-like components in these cells. 6. Glycoproteins of MCF-7 cells interacting with anti-glycophorin antibodies were affinity purified and partially characterized.
Barsoum
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AL
Department of Biological Chemistry, Milton S. Hershey Medical Center, PA 17033.
Bhavanandan
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VP
D001943:Breast Neoplasms / Q000032:analysis; D002460:Cell Line; D002478:Cells, Cultured; D002846:Chromatography, Affinity; D004910:Erythrocyte Membrane / Q000032:analysis*; D005455:Fluorescent Antibody Technique; D006021:Glycophorins / Q000032:analysis* / Q000302:isolation & purification; D006801:Humans; D007106:Immune Sera / Q000032:analysis; D005779:Immunodiffusion; D008562:Membrane Glycoproteins / Q000097:blood* / Q000302:isolation & purification; D012685:Sepharose / Q000031:analogs & derivatives; D012795:Sialoglycoproteins / Q000032:analysis*
D006021:Glycophorins; D007106:Immune Sera; D008562:Membrane Glycoproteins; D012795:Sialoglycoproteins; C036210:immunoglobulin G-sepharose; D012685:Sepharose