Donnell M E, DeLuca G M, Blackall D P
Department of Pathology, University of Tennessee College of Medicine, Memphis 38163, USA.
J Immunol Methods. 1995 Nov 16;187(1):111-9. doi: 10.1016/0022-1759(95)00175-a.
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.
质膜囊泡已被用于研究多种生物学事件，包括受体 - 配体相互作用的性质以及分子跨膜运输的生理学。质膜囊泡是从表达重组血型糖蛋白A（一种研究充分的红细胞内在膜蛋白）的贴壁中国仓鼠卵巢细胞系中化学诱导产生的。它们也从悬浮培养的中国仓鼠卵巢细胞中制备。生化和免疫分析表明细胞膜和囊泡上的血型糖蛋白A具有等效性。转染的中国仓鼠卵巢细胞膜也用一种高度脂肪族荧光细胞连接剂进行了标记。由荧光素标记的细胞产生的囊泡显示出质膜的明亮表面染色。它们在生化和免疫方面也表达与基于细胞的血型糖蛋白A无法区分的血型糖蛋白A。质膜囊泡在培养中不贴壁，并在细胞膜中稳定保留荧光探针。荧光素标记的囊泡对于研究受体 - 配体对的两种成分都在贴壁细胞系上表达的细胞相互作用将特别有用。未标记的囊泡在临床实验室和基础科学研究环境中作为可溶性免疫吸附剂也可能被证明是有用的。
