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*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

719 Scopus citations

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

Funding

FundersFunder number
Fulbright Foundation
National Institutes of Health
National Institute of General Medical SciencesR01GM036548

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