Direct demonstration of the endocytic function of caveolae by a cell-free assay

Article Article scientifique Article Direct demonstration of the endocytic function of caveolae by a cell-free assay eng 1999 author Gilbert Anne 27312 author Paccaud J P author Foti Michelangelo 0000-0001-7199-4135 28033 author Porcheron Geneviève 26280 author Balz J author Carpentier J L 1101-1110 10.1242/jcs.112.7.1101 10198292 0021-9533 The endocytic function of caveolae was challenged by taking advantage of a cell-free assay directly measuring the detachment of receptor-containing vesicles from isolated plasma membranes. Plasma membranes from cultured cells surface-labeled with 125I-cholera toxin (segregating in caveolae) were isolated as described previously. Following incubation of these labeled membranes in the presence of nucleotide(s) and cytosol, a significant proportion of the initially membrane-associated radioactivity was released into the incubation medium in sedimentable form (14*10(6 )g). Results of biochemical, morphological, and fractionation analysis of the material containing the released radioactivity directly demonstrated that caveolae are plasma membrane domains involved in an endocytic process and resulting in the formation of caveolae-derived vesicles. In addition, these studies allowed a direct comparison of caveolae- and clathrin-coated pit-mediated endocytosis and reveal that these two processes diverge in terms of kinetics, cytosol and nucleotide requirements as well as in terms of the density and size of the endocytic vesicles formed. 3T3 Cells Adenosine Triphosphate/metabolism Animals Cell Membrane/physiology/ultrastructure Cell-Free System/metabolism Centrifugation Density Gradient Coated Pits Cell-Membrane/physiology Cytosol/metabolism Edetic Acid/pharmacology Endocytosis/physiology Filipin/pharmacology Guanosine Triphosphate/metabolism Mice Microscopy Electron Radioligand Assay/methods Time Factors Dewey 612 134 Département de physiologie cellulaire et métabolisme 21C false 472 Obésité et syndrome métabolique 803 Journal of cell science 112 ( Pt 7) 4893521d-ee70-48c6-8551-1325073481a7 Article (Published version) Restricted Gilbert A JCS 1999.pdf 388286 application/pdf restrictedAccess Faculté de médecine / Section de médecine fondamentale / Département de physiologie cellulaire et métabolisme

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Direct demonstration of the endocytic function of caveolae by a cell-free assay	Journal of cell science			1999	408	0

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Gilbert, Anne