Endocytosis by random initiation and stabilization of clathrin-coated pits

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

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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.

Original languageEnglish (US)
Pages (from-to)591-605
Number of pages15
JournalCell
Volume118
Issue number5
DOIs
StatePublished - Sep 3 2004

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ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Ehrlich, M., Boll, W., Van Oijen, A., Hariharan, R., Chandran, K., Nibert, M. L., & Kirchhausen, T. (2004). Endocytosis by random initiation and stabilization of clathrin-coated pits. Cell, 118(5), 591-605. https://doi.org/10.1016/j.cell.2004.08.017