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. 2004 Sep 3;118(5):591-605.
doi: 10.1016/j.cell.2004.08.017.

Endocytosis by random initiation and stabilization of clathrin-coated pits

Marcelo Ehrlich  1 , Werner Boll, Antoine Van Oijen, Ramesh Hariharan, Kartik Chandran, Max L Nibert, Tomas Kirchhausen
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Free article

Endocytosis by random initiation and stabilization of clathrin-coated pits

Marcelo Ehrlich et al. Cell. .
Free article

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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