TY - JOUR TI - Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions AU - Thompson, Connor J AU - Su, Zhaoqian AU - Vu, Vinh H AU - Wu, Yinghao AU - Leckband, Deborah E AU - Schwartz, Daniel K A2 - Ben-Tal, Nir A2 - Boudker, Olga VL - 9 PY - 2020 DA - 2020/09/02 SP - e59035 C1 - eLife 2020;9:e59035 DO - 10.7554/eLife.59035 UR - https://doi.org/10.7554/eLife.59035 AB - 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. KW - E-cadherin KW - membrane protein KW - lateral interactions KW - single-molecule KW - adherens junctions KW - cell adhesion JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -