Cytosol- and clathrin-dependent stimulation of endocytosis in vitro by purified adaptors

Elizabeth Smythe, Laura L. Carter, Sandra L. Schmid

Research output: Contribution to journalArticle

65 Scopus citations

Abstract

Using stage-specific assays for receptor-mediated endocytosis of transferrin (Tfn) into perforated A431 cells we show that purified adaptors stimulate coated pit assembly and ligand sequestration into deeply invaginated coated pits. Late events in endocytosis involving membrane fission and coated vesicle budding which lead to the internalization of Tfn are unaffected. AP2, plasma membrane adaptors, are active at physiological concentrations, whereas API, Golgi adaptors, are inactive. Adaptor-dependent stimulation of Tfn sequestration requires cytosolic clathrin, but is unaffected by clathrin purified from coated vesicles suggesting that soluble and assembled clathrin pools are functionally distinct. In addition to adaptors and cytosolic clathrin other, as yet unidentified, cytosolic factors are also required for efficient coated pit invagination. These results provide new insight into the mechanisms and regulation of coated pit assembly and invagination.

Original languageEnglish (US)
Pages (from-to)1163-1171
Number of pages9
JournalJournal of Cell Biology
Volume119
Issue number5
DOIs
StatePublished - Dec 1992

ASJC Scopus subject areas

  • Cell Biology

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