Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions
- University of Colorado Boulder, United States
- Albert Einstein College of Medicine, United States
- University of Illinois, Urbana−Champaign, United States
- 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
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.
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.
Metrics
-
- 2,435
- views
-
- 264
- downloads
-
- 36
- citations
Views, downloads and citations are aggregated across all versions of this paper published by eLife.
Download links
A two-part list of links to download the article, or parts of the article, in various formats.
Downloads (link to download the article as PDF)
Open citations (links to open the citations from this article in various online reference manager services)
Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)
Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions
eLife 9:e59035.
https://doi.org/10.7554/eLife.59035
