Preparation and characterization of fluorescein-labeled plasma membrane vesicles from transfected Chinese hamster ovary cells.
Plasma membrane vesicles have been used to study a wide variety of biological events including the nature of receptor-ligand interactions and the physiology of molecular transport across membranes. Plasma membrane vesicles were chemically induced from an adherent Chinese hamster ovary cell line expressing recombinant glycophorin A, a well-studied intrinsic membrane protein of the red blood cell. They were also prepared from Chinese hamster ovary cells in suspension culture. Biochemical and immunological assays demonstrated the equivalence of glycophorin A on cell membranes and vesicles. The transfected Chinese hamster ovary cell membrane was also labeled with a highly aliphatic fluorescent cell linker. Vesicles produced by the fluorescein-labeled cells demonstrated bright surface staining of the plasma membrane. They too expressed glycophorin A biochemically and immunologically indistinguishable from cellular-based glycophorin A. Plasma membrane vesicles are non-adherent in culture and stably retain the fluorescent probe in the cell membrane. Fluorescein-labeled vesicles will be particularly useful for studying cellular interactions in which both constituents of a receptor-ligand pair are expressed on adherent cell lines. Unlabeled vesicles may also prove to be useful as soluble immunoadsorbants in both the clinical laboratory and basic science research settings.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D015153:Blotting, Western; D016466:CHO Cells; D002462:Cell Membrane / Q000276:immunology*; D006224:Cricetinae; D019793:Fluorescein; D005452:Fluoresceins; D006021:Glycophorins / Q000032:analysis*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008861:Microsomes / Q000276:immunology; D014162:Transfection
D000911:Antibodies, Monoclonal; D005452:Fluoresceins; D006021:Glycophorins; D019793:Fluorescein