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Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin
The advent of super-resolution imaging (SRI) has created a need for optimized labelling strategies. We present a new method relying on fluorophore-conjugated monomeric streptavidin (mSA) to label membrane proteins carrying a short, enzymatically biotinylated tag, compatible with SRI techniques inclu...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4799371/ https://www.ncbi.nlm.nih.gov/pubmed/26979420 http://dx.doi.org/10.1038/ncomms10773 |
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author | Chamma, Ingrid Letellier, Mathieu Butler, Corey Tessier, Béatrice Lim, Kok-Hong Gauthereau, Isabel Choquet, Daniel Sibarita, Jean-Baptiste Park, Sheldon Sainlos, Matthieu Thoumine, Olivier |
author_facet | Chamma, Ingrid Letellier, Mathieu Butler, Corey Tessier, Béatrice Lim, Kok-Hong Gauthereau, Isabel Choquet, Daniel Sibarita, Jean-Baptiste Park, Sheldon Sainlos, Matthieu Thoumine, Olivier |
author_sort | Chamma, Ingrid |
collection | PubMed |
description | The advent of super-resolution imaging (SRI) has created a need for optimized labelling strategies. We present a new method relying on fluorophore-conjugated monomeric streptavidin (mSA) to label membrane proteins carrying a short, enzymatically biotinylated tag, compatible with SRI techniques including uPAINT, STED and dSTORM. We demonstrate efficient and specific labelling of target proteins in confined intercellular and organotypic tissues, with reduced steric hindrance and no crosslinking compared with multivalent probes. We use mSA to decipher the dynamics and nanoscale organization of the synaptic adhesion molecules neurexin-1β, neuroligin-1 (Nlg1) and leucine-rich-repeat transmembrane protein 2 (LRRTM2) in a dual-colour configuration with GFP nanobody, and show that these proteins are diffusionally trapped at synapses where they form apposed trans-synaptic adhesive structures. Furthermore, Nlg1 is dynamic, disperse and sensitive to synaptic stimulation, whereas LRRTM2 is organized in compact and stable nanodomains. Thus, mSA is a versatile tool to image membrane proteins at high resolution in complex live environments, providing novel information about the nano-organization of biological structures. |
format | Online Article Text |
id | pubmed-4799371 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-47993712016-03-23 Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin Chamma, Ingrid Letellier, Mathieu Butler, Corey Tessier, Béatrice Lim, Kok-Hong Gauthereau, Isabel Choquet, Daniel Sibarita, Jean-Baptiste Park, Sheldon Sainlos, Matthieu Thoumine, Olivier Nat Commun Article The advent of super-resolution imaging (SRI) has created a need for optimized labelling strategies. We present a new method relying on fluorophore-conjugated monomeric streptavidin (mSA) to label membrane proteins carrying a short, enzymatically biotinylated tag, compatible with SRI techniques including uPAINT, STED and dSTORM. We demonstrate efficient and specific labelling of target proteins in confined intercellular and organotypic tissues, with reduced steric hindrance and no crosslinking compared with multivalent probes. We use mSA to decipher the dynamics and nanoscale organization of the synaptic adhesion molecules neurexin-1β, neuroligin-1 (Nlg1) and leucine-rich-repeat transmembrane protein 2 (LRRTM2) in a dual-colour configuration with GFP nanobody, and show that these proteins are diffusionally trapped at synapses where they form apposed trans-synaptic adhesive structures. Furthermore, Nlg1 is dynamic, disperse and sensitive to synaptic stimulation, whereas LRRTM2 is organized in compact and stable nanodomains. Thus, mSA is a versatile tool to image membrane proteins at high resolution in complex live environments, providing novel information about the nano-organization of biological structures. Nature Publishing Group 2016-03-16 /pmc/articles/PMC4799371/ /pubmed/26979420 http://dx.doi.org/10.1038/ncomms10773 Text en Copyright © 2016, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved. http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Chamma, Ingrid Letellier, Mathieu Butler, Corey Tessier, Béatrice Lim, Kok-Hong Gauthereau, Isabel Choquet, Daniel Sibarita, Jean-Baptiste Park, Sheldon Sainlos, Matthieu Thoumine, Olivier Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
title | Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
title_full | Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
title_fullStr | Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
title_full_unstemmed | Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
title_short | Mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
title_sort | mapping the dynamics and nanoscale organization of synaptic adhesion proteins using monomeric streptavidin |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4799371/ https://www.ncbi.nlm.nih.gov/pubmed/26979420 http://dx.doi.org/10.1038/ncomms10773 |
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