Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions

  1. Connor J Thompson
  2. Zhaoqian Su
  3. Vinh H Vu
  4. Yinghao Wu
  5. Deborah E Leckband
  6. Daniel K Schwartz  Is a corresponding author
  1. University of Colorado Boulder, United States
  2. Albert Einstein College of Medicine, United States
  3. University of Illinois, Urbana−Champaign, United States
  4. University of Illinois, United States

Abstract

We demonstrate a combined experimental and computational approach for the quantitative characterization of lateral interactions between membrane-associated proteins. In particular, weak, lateral (cis) interactions between E-cadherin extracellular domains tethered to supported lipid bilayers, were studied using a combination of dynamic single-molecule Förster Resonance Energy Transfer (FRET) and kinetic Monte Carlo (kMC) simulations. Cadherins are intercellular adhesion proteins that assemble into clusters at cell-cell contacts through cis- and trans- (adhesive) interactions. A detailed and quantitative understanding of cis-clustering has been hindered by a lack of experimental approaches capable of detecting and quantifying lateral interactions between proteins on membranes. Here single-molecule intermolecular FRET measurements of wild-type E-cadherin and cis-interaction mutants combined with simulations demonstrate that both nonspecific and specific cis-interactions contribute to lateral clustering on lipid bilayers. Moreover, the intermolecular binding and dissociation rate constants are quantitatively and independently determined, demonstrating an approach that is generalizable for other interacting proteins.

Data availability

All data generated or analyzed in this work are included in the main text, figure supplements, and Supplementary File 1.

Article and author information

Author details

  1. Connor J Thompson

    Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6226-7171
  2. Zhaoqian Su

    Department of Systems & Computational Biology, Albert Einstein College of Medicine, The Bronx, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8369-0697
  3. Vinh H Vu

    Department of Biochemistry, University of Illinois, Urbana−Champaign, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  4. Yinghao Wu

    Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, United States
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1181-5670
  5. Deborah E Leckband

    Chemistry and Chemical and Biomolecular Engineering, University of Illinois, Urbana, United States
    Competing interests
    The authors declare that no competing interests exist.
  6. Daniel K Schwartz

    Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, United States
    For correspondence
    Daniel.schwartz@colorado.edu
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-5397-7200

Funding

National Institute of General Medical Sciences (1R01GM117104)

  • Connor J Thompson
  • Zhaoqian Su
  • Vinh H Vu
  • Yinghao Wu
  • Deborah E Leckband
  • Daniel K Schwartz

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Nir Ben-Tal, Tel Aviv University, Israel

Version history

  1. Received: May 18, 2020
  2. Accepted: September 1, 2020
  3. Accepted Manuscript published: September 2, 2020 (version 1)
  4. Version of Record published: September 21, 2020 (version 2)

Copyright

© 2020, Thompson et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Connor J Thompson
  2. Zhaoqian Su
  3. Vinh H Vu
  4. Yinghao Wu
  5. Deborah E Leckband
  6. Daniel K Schwartz
(2020)
Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions
eLife 9:e59035.
https://doi.org/10.7554/eLife.59035

Share this article

https://doi.org/10.7554/eLife.59035

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