Interactome Analysis Reveals Ezrin Can Adopt Multiple Conformational States*

Background: Ezrin is a conformationally regulated microfilament-membrane linker restricted to the apical aspect of epithelial cells. Results: Quantitative mass spectrometry identified proteins that associate with different ezrin conformations; the interactors include novel apical microvilli-associated proteins, with classes perceiving ezrin's conformation differentially. Conclusion: Different proteins selectively associate with three distinct conformational states of ezrin. Significance: This study extends the conformational activation model of ezrin and identifies new interacting partners. Ezrin, a member of the ezrin-radixin-moesin family (ERM), is an essential regulator of the structure of microvilli on the apical aspect of epithelial cells. Ezrin provides a linkage between membrane-associated proteins and F-actin, oscillating between active/open and inactive/closed states, and is regulated in part by phosphorylation of a C-terminal threonine. In the open state, ezrin can bind a number of ligands, but in the closed state the ligand-binding sites are inaccessible. In vitro analysis has proposed that there may be a third hyperactivated form of ezrin. To gain a better understanding of ezrin, we conducted an unbiased proteomic analysis of ezrin-binding proteins in an epithelial cell line, Jeg-3. We refined our list of interactors by comparing the interactomes using quantitative mass spectrometry between wild-type ezrin, closed ezrin, open ezrin, and hyperactivated ezrin. The analysis reveals several novel interactors confirmed by their localization to microvilli, as well as a significant class of proteins that bind closed ezrin. Taken together, the data indicate that ezrin can exist in three different conformational states, and different ligands “perceive” ezrin conformational states differently.

• Publication year

  2013

• Venue

  Journal of Biological Chemistry

• Publication date

  2013-10-22

• Fields of study

  Biology, Medicine

• Identifiers
  DOI 10.1074/jbc.M113.505669PMID 24151071PMCID PMC3853291
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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