ATP- and cytosol-dependent release of adaptor proteins from clathrin- coated vesicles: A dual role for hsc70

Lisa A. Hannan, Sherri L. Newmyer, Sandra L. Schmid

Research output: Contribution to journalArticle

56 Citations (Scopus)

Abstract

Clathrin-coated vesicles (CCV) mediate protein sorting and vesicular trafficking from the plasma membrane and the trans-Golgi network. Before delivery of the vesicle contents to the target organelles, the coat components, clathrin and adaptor protein complexes (APs), must be released. Previous work has established that hsc70/the uncoating ATPase mediates clathrin release in vitro without the release of APs. AP release has not been reconstituted in vitro, and nothing is known about the requirements for this reaction. We report a novel quantitative assay for the ATP- and cytosol- dependent release of APs from CCV. As expected, hsc70 is not sufficient for AP release; however, immunodepletion and reconstitution experiments establish that it is necessary. Interestingly, complete clathrin release is not a prerequisite for AP release, suggesting that hsc70 plays a dual role in recycling the constituents of the clathrin coat. This assay provides a functional basis for identification of the additional cytosolic factor(s) required for AP release.

Original languageEnglish (US)
Pages (from-to)2217-2229
Number of pages13
JournalMolecular Biology of the Cell
Volume9
Issue number8
StatePublished - Aug 1998

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Clathrin-Coated Vesicles
Clathrin
Cytosol
Adenosine Triphosphate
Vesicular Transport Adaptor Proteins
trans-Golgi Network
Proteins
Recycling
Protein Transport
Organelles
Adenosine Triphosphatases
Cell Membrane
In Vitro Techniques

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

Cite this

ATP- and cytosol-dependent release of adaptor proteins from clathrin- coated vesicles : A dual role for hsc70. / Hannan, Lisa A.; Newmyer, Sherri L.; Schmid, Sandra L.

In: Molecular Biology of the Cell, Vol. 9, No. 8, 08.1998, p. 2217-2229.

Research output: Contribution to journalArticle

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