EHD2 restrains dynamics of caveolae by an ATP-dependent, membrane-bound, open conformation

Proc Natl Acad Sci U S A. 2017 May 30;114(22):E4360-E4369. doi: 10.1073/pnas.1614066114. Epub 2017 Feb 21.


The EH-domain-containing protein 2 (EHD2) is a dynamin-related ATPase that confines caveolae to the cell surface by restricting the scission and subsequent endocytosis of these membrane pits. For this, EHD2 is thought to first bind to the membrane, then to oligomerize, and finally to detach, in a stringently regulated mechanistic cycle. It is still unclear how ATP is used in this process and whether membrane binding is coupled to conformational changes in the protein. Here, we show that the regulatory N-terminal residues and the EH domain keep the EHD2 dimer in an autoinhibited conformation in solution. By significantly advancing the use of infrared reflection-absorption spectroscopy, we demonstrate that EHD2 adopts an open conformation by tilting the helical domains upon membrane binding. We show that ATP binding enables partial insertion of EHD2 into the membrane, where G-domain-mediated oligomerization occurs. ATP hydrolysis is related to detachment of EHD2 from the membrane. Finally, we demonstrate that the regulation of EHD2 oligomerization in a membrane-bound state is crucial to restrict caveolae dynamics in cells.

Keywords: EHD2; caveolae; infrared reflection–absorption spectroscopy; membrane-bound protein structure; membrane-reshaping protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Amino Acid Substitution
  • Animals
  • Carrier Proteins / chemistry*
  • Carrier Proteins / genetics
  • Carrier Proteins / metabolism*
  • Caveolae / metabolism
  • Fluorescence Resonance Energy Transfer
  • HeLa Cells
  • Humans
  • Mice
  • Molecular Dynamics Simulation
  • Mutagenesis, Site-Directed
  • Protein Binding
  • Protein Conformation
  • Protein Structure, Quaternary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Spectrophotometry, Infrared


  • Carrier Proteins
  • EHD2 protein, mouse
  • Recombinant Proteins
  • Adenosine Triphosphate