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Single-molecule force spectroscopy of protein-membrane interactions
Many biological processes rely on protein–membrane interactions in the presence of mechanical forces, yet high resolution methods to quantify such interactions are lacking. Here, we describe a single-molecule force spectroscopy approach to quantify membrane binding of C2 domains in Synaptotagmin-1 (...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5690283/ https://www.ncbi.nlm.nih.gov/pubmed/29083305 http://dx.doi.org/10.7554/eLife.30493 |
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author | Ma, Lu Cai, Yiying Li, Yanghui Jiao, Junyi Wu, Zhenyong O'Shaughnessy, Ben De Camilli, Pietro Karatekin, Erdem Zhang, Yongli |
author_facet | Ma, Lu Cai, Yiying Li, Yanghui Jiao, Junyi Wu, Zhenyong O'Shaughnessy, Ben De Camilli, Pietro Karatekin, Erdem Zhang, Yongli |
author_sort | Ma, Lu |
collection | PubMed |
description | Many biological processes rely on protein–membrane interactions in the presence of mechanical forces, yet high resolution methods to quantify such interactions are lacking. Here, we describe a single-molecule force spectroscopy approach to quantify membrane binding of C2 domains in Synaptotagmin-1 (Syt1) and Extended Synaptotagmin-2 (E-Syt2). Syts and E-Syts bind the plasma membrane via multiple C2 domains, bridging the plasma membrane with synaptic vesicles or endoplasmic reticulum to regulate membrane fusion or lipid exchange, respectively. In our approach, single proteins attached to membranes supported on silica beads are pulled by optical tweezers, allowing membrane binding and unbinding transitions to be measured with unprecedented spatiotemporal resolution. C2 domains from either protein resisted unbinding forces of 2–7 pN and had binding energies of 4–14 k(B)T per C2 domain. Regulation by bilayer composition or Ca(2+) recapitulated known properties of both proteins. The method can be widely applied to study protein–membrane interactions. |
format | Online Article Text |
id | pubmed-5690283 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-56902832017-11-20 Single-molecule force spectroscopy of protein-membrane interactions Ma, Lu Cai, Yiying Li, Yanghui Jiao, Junyi Wu, Zhenyong O'Shaughnessy, Ben De Camilli, Pietro Karatekin, Erdem Zhang, Yongli eLife Structural Biology and Molecular Biophysics Many biological processes rely on protein–membrane interactions in the presence of mechanical forces, yet high resolution methods to quantify such interactions are lacking. Here, we describe a single-molecule force spectroscopy approach to quantify membrane binding of C2 domains in Synaptotagmin-1 (Syt1) and Extended Synaptotagmin-2 (E-Syt2). Syts and E-Syts bind the plasma membrane via multiple C2 domains, bridging the plasma membrane with synaptic vesicles or endoplasmic reticulum to regulate membrane fusion or lipid exchange, respectively. In our approach, single proteins attached to membranes supported on silica beads are pulled by optical tweezers, allowing membrane binding and unbinding transitions to be measured with unprecedented spatiotemporal resolution. C2 domains from either protein resisted unbinding forces of 2–7 pN and had binding energies of 4–14 k(B)T per C2 domain. Regulation by bilayer composition or Ca(2+) recapitulated known properties of both proteins. The method can be widely applied to study protein–membrane interactions. eLife Sciences Publications, Ltd 2017-10-30 /pmc/articles/PMC5690283/ /pubmed/29083305 http://dx.doi.org/10.7554/eLife.30493 Text en © 2017, Ma 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 Ma, Lu Cai, Yiying Li, Yanghui Jiao, Junyi Wu, Zhenyong O'Shaughnessy, Ben De Camilli, Pietro Karatekin, Erdem Zhang, Yongli Single-molecule force spectroscopy of protein-membrane interactions |
title | Single-molecule force spectroscopy of protein-membrane interactions |
title_full | Single-molecule force spectroscopy of protein-membrane interactions |
title_fullStr | Single-molecule force spectroscopy of protein-membrane interactions |
title_full_unstemmed | Single-molecule force spectroscopy of protein-membrane interactions |
title_short | Single-molecule force spectroscopy of protein-membrane interactions |
title_sort | single-molecule force spectroscopy of protein-membrane interactions |
topic | Structural Biology and Molecular Biophysics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5690283/ https://www.ncbi.nlm.nih.gov/pubmed/29083305 http://dx.doi.org/10.7554/eLife.30493 |
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