Insulin-sensitive association of GLUT-4 with endocytic clathrin-coated vesicles revealed with the use of brefeldin A.
作者： R. Chakrabarti , J. Buxton , M. Joly , S. Corvera
DOI: 10.1016/S0021-9258(17)37140-5
关键词:
摘要: The interaction of the adipocyte/skeletal muscle glucose transporter (GLUT-4) with clathrin lattices may be important in maintaining its intracellular distribution. To better understand role sorting GLUT-4, we have attempted to determine cellular origin clathrin-coated vesicles (CCVs) that contain this transporter. fungal toxin brefeldin A (BFA) causes selective disassembly at trans-Golgi network (TGN), but not plasma membrane (PM), thus providing a way estimating proportion GLUT-4 PM- versus TGN-derived lattices. Exposure 3T3-L1 adipocytes BFA resulted rapid TGN, observed by optical sectioning microscopy, and pronounced decrease yield CCVs purified from these cells. Thus, isolated BFA-treated cells are likely derived PM. Immunoblotting experiments revealed presence such CCVs, suggesting under basal conditions is continually retrieved PM through CCV pathway. both or non-treated insulin 4-6-fold increase concentration In parallel, PM-derived decreased 60%. These results suggest (a) effect cell surface inhibited BFA, (b) association endocytic facilitating increased response hormone.
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