The AP2 adaptor enhances clathrin coat stiffness

Michael Lherbette, Lisa Redlingshöfer, Frances M. Brodsky, Iwan A. T. Schaap, Philip N. Dannhauser

Research output: Contribution to journalArticle

5 Citations (Scopus)
4 Downloads (Pure)

Abstract

Deformation of the plasma membrane into clathrin-coated vesicles is a critical step in clathrin-mediated endocytosis and requires the orchestrated assembly of clathrin and endocytic adaptors into a membrane-associated protein coat. The individual role of these membrane-bending and curvature-stabilizing factors is subject to current debate. As such, it is unclear whether the clathrin coat itself is stiff enough to impose curvature and if so, whether this could be effectively transferred to the membrane by the linking adaptor proteins. We have recently demonstrated that clathrin alone is sufficient to form membrane buds in vitro. Here, we use atomic force microscopy to assess the contributions of clathrin and its membrane adaptor protein 2 (AP2) to clathrin coat stiffness, which determines the mechanics of vesicle formation. We found that clathrin coats are less than 10-fold stiffer than the membrane they enclose, suggesting a delicate balance between the forces harnessed from clathrin coat formation and those required for membrane bending. We observed that clathrin adaptor protein AP2 increased the stiffness of coats formed from native clathrin, but did not affect less-flexible coats formed from clathrin lacking the light chain subunits. We thus propose that clathrin light chains are important for clathrin coat flexibility and that AP2 facilitates efficient cargo sequestration during coated vesicle formation by modulating clathrin coat stiffness.

Original languageEnglish
Pages (from-to)4074-4085
Number of pages12
JournalFEBS Journal
Volume286
Issue number20
Early online date14 Jun 2019
DOIs
Publication statusPublished - Oct 2019

Keywords

  • AFM
  • clathrin
  • membrane biophysics

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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