Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions
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 st...
| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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eLife Sciences Publications Ltd
2020-09-01
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| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/59035 |
| _version_ | 1811180937121103872 |
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| author | Connor J Thompson Zhaoqian Su Vinh H Vu Yinghao Wu Deborah E Leckband Daniel K Schwartz |
| author_facet | Connor J Thompson Zhaoqian Su Vinh H Vu Yinghao Wu Deborah E Leckband Daniel K Schwartz |
| author_sort | Connor J Thompson |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-04-11T09:11:42Z |
| format | Article |
| id | doaj.art-5bb1ac7860544a73ac2e02b89448891b |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-11T09:11:42Z |
| publishDate | 2020-09-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-5bb1ac7860544a73ac2e02b89448891b2022-12-22T04:32:29ZengeLife Sciences Publications LtdeLife2050-084X2020-09-01910.7554/eLife.59035Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactionsConnor J Thompson0https://orcid.org/0000-0001-6226-7171Zhaoqian Su1https://orcid.org/0000-0002-8369-0697Vinh H Vu2Yinghao Wu3https://orcid.org/0000-0003-1181-5670Deborah E Leckband4Daniel K Schwartz5https://orcid.org/0000-0001-5397-7200Department of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, United StatesDepartment of Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, United StatesDepartment of Biochemistry and University of Illinois, Urbana-Champaign, Urbana, United StatesDepartment of Systems and Computational Biology, Albert Einstein College of Medicine, Bronx, United StatesDepartment of Biochemistry and University of Illinois, Urbana-Champaign, Urbana, United States; Department of Chemical and Biomolecular Engineering, University of Illinois, Urbana-Champaign, Urbana, United StatesDepartment of Chemical and Biological Engineering, University of Colorado Boulder, Boulder, United StatesWe 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.https://elifesciences.org/articles/59035E-cadherinmembrane proteinlateral interactionssingle-moleculeadherens junctionscell adhesion |
| spellingShingle | Connor J Thompson Zhaoqian Su Vinh H Vu Yinghao Wu Deborah E Leckband Daniel K Schwartz Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions eLife E-cadherin membrane protein lateral interactions single-molecule adherens junctions cell adhesion |
| title | Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions |
| title_full | Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions |
| title_fullStr | Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions |
| title_full_unstemmed | Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions |
| title_short | Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions |
| title_sort | cadherin clusters stabilized by a combination of specific and nonspecific cis interactions |
| topic | E-cadherin membrane protein lateral interactions single-molecule adherens junctions cell adhesion |
| url | https://elifesciences.org/articles/59035 |
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