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      Endocytosis by random initiation and stabilization of clathrin-coated pits.

      Cell

      Adaptor Protein Complex 2, metabolism, Animals, Cell Line, Cell Membrane, ultrastructure, Clathrin-Coated Vesicles, Endocytosis, physiology, Endosomes, Green Fluorescent Proteins, Haplorhini, Lipoproteins, LDL, Luminescent Proteins, Models, Biological, Protein Transport, Rats, Recombinant Fusion Proteins, Reoviridae, Transferrin

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          Abstract

          Clathrin-coated vesicles carry traffic from the plasma membrane to endosomes. We report here the real-time visualization of cargo sorting and endocytosis by clathrin-coated pits in living cells. We have detected the formation of coats by monitoring incorporation of fluorescently tagged clathrin or its adaptor AP-2; we have also followed clathrin-mediated uptake of transferrin and of single LDL or reovirus particles. The intensity of a cargo-loaded clathrin cluster grows steadily during its lifetime, and the time required to complete assembly is proportional to the size of the cargo particle. These results are consistent with a nucleation-growth mechanism and an approximately constant growth rate. There are no strongly preferred nucleation sites. A proportion of the nucleation events are weak and short lived. Cargo incorporation occurs primarily or exclusively in a newly formed coated pit. Our data lead to a model in which coated pits initiate randomly but collapse unless stabilized, perhaps by cargo capture.

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          Journal
          15339664
          10.1016/j.cell.2004.08.017

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