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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...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7505656/ https://www.ncbi.nlm.nih.gov/pubmed/32876051 http://dx.doi.org/10.7554/eLife.59035 |
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author | Thompson, Connor J Su, Zhaoqian Vu, Vinh H Wu, Yinghao Leckband, Deborah E Schwartz, Daniel K |
author_facet | Thompson, Connor J Su, Zhaoqian Vu, Vinh H Wu, Yinghao Leckband, Deborah E Schwartz, Daniel K |
author_sort | Thompson, Connor J |
collection | PubMed |
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. |
format | Online Article Text |
id | pubmed-7505656 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-75056562020-09-23 Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions Thompson, Connor J Su, Zhaoqian Vu, Vinh H Wu, Yinghao Leckband, Deborah E Schwartz, Daniel K eLife Structural Biology and Molecular Biophysics 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. eLife Sciences Publications, Ltd 2020-09-02 /pmc/articles/PMC7505656/ /pubmed/32876051 http://dx.doi.org/10.7554/eLife.59035 Text en © 2020, Thompson et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Structural Biology and Molecular Biophysics Thompson, Connor J Su, Zhaoqian Vu, Vinh H Wu, Yinghao Leckband, Deborah E Schwartz, Daniel K Cadherin clusters stabilized by a combination of specific and nonspecific cis-interactions |
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 | Structural Biology and Molecular Biophysics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7505656/ https://www.ncbi.nlm.nih.gov/pubmed/32876051 http://dx.doi.org/10.7554/eLife.59035 |
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