Understanding Living Clathrin-Coated Pits

Authors

  • Joshua Z. Rappoport,

    1. The Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, Box 304, New York, New York 10021, USA
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  • Sanford M. Simon,

    1. The Laboratory of Cellular Biophysics, The Rockefeller University, 1230 York Avenue, Box 304, New York, New York 10021, USA
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  • Alexandre Benmerah

    Corresponding author
    1. Department of Infectious Diseases, Institut Cochin (INSERM U567, CNRS UMR 8104, Universitè Paris 5), 27 rue du Faubourg St Jacques, 75014 Paris, France
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A. Benmerah, benmerah@cochin.inserm.fr;S.Simon,Simon@Rockefeller.edu

Abstract

Most knowledge of clathrin-mediated endocytosis has been gained by biochemical fractionation and in vitro assays. Recently, the study of endocytosis has extended into the living cell. The tracking of individual clathrin-coated pits and vesicles (CCPs and CCVs) has provided new insight into understanding the dynamic nature of CCPs. The use of total internal reflection fluorescence microscopy (TIR-FM), also termed evanescent field microscopy, has enabled the direct observation of events occurring within a restricted area of the cell adjacent to and including the adherent plasma membrane. TIR-FM is now actively being pursued in the study of endocytic processes. The direct observation of CCP-associated proteins including clathrin itself, dynamin and, most recently, AP-2 has considerably challenged old models, confirming some points but raising very interesting new questions.

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