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Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae

In mammalian cells three closely related cavin proteins cooperate with the scaffolding protein caveolin to form membrane invaginations known as caveolae. Here we have developed a novel single-molecule fluorescence approach to directly observe interactions and stoichiometries in protein complexes fro...

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Autores principales: Gambin, Yann, Ariotti, Nicholas, McMahon, Kerrie-Ann, Bastiani, Michele, Sierecki, Emma, Kovtun, Oleksiy, Polinkovsky, Mark E, Magenau, Astrid, Jung, WooRam, Okano, Satomi, Zhou, Yong, Leneva, Natalya, Mureev, Sergey, Johnston, Wayne, Gaus, Katharina, Hancock, John F, Collins, Brett M, Alexandrov, Kirill, Parton, Robert G
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3903133/
https://www.ncbi.nlm.nih.gov/pubmed/24473072
http://dx.doi.org/10.7554/eLife.01434
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author Gambin, Yann
Ariotti, Nicholas
McMahon, Kerrie-Ann
Bastiani, Michele
Sierecki, Emma
Kovtun, Oleksiy
Polinkovsky, Mark E
Magenau, Astrid
Jung, WooRam
Okano, Satomi
Zhou, Yong
Leneva, Natalya
Mureev, Sergey
Johnston, Wayne
Gaus, Katharina
Hancock, John F
Collins, Brett M
Alexandrov, Kirill
Parton, Robert G
author_facet Gambin, Yann
Ariotti, Nicholas
McMahon, Kerrie-Ann
Bastiani, Michele
Sierecki, Emma
Kovtun, Oleksiy
Polinkovsky, Mark E
Magenau, Astrid
Jung, WooRam
Okano, Satomi
Zhou, Yong
Leneva, Natalya
Mureev, Sergey
Johnston, Wayne
Gaus, Katharina
Hancock, John F
Collins, Brett M
Alexandrov, Kirill
Parton, Robert G
author_sort Gambin, Yann
collection PubMed
description In mammalian cells three closely related cavin proteins cooperate with the scaffolding protein caveolin to form membrane invaginations known as caveolae. Here we have developed a novel single-molecule fluorescence approach to directly observe interactions and stoichiometries in protein complexes from cell extracts and from in vitro synthesized components. We show that up to 50 cavins associate on a caveola. However, rather than forming a single coat complex containing the three cavin family members, single-molecule analysis reveals an exquisite specificity of interactions between cavin1, cavin2 and cavin3. Changes in membrane tension can flatten the caveolae, causing the release of the cavin coat and its disassembly into separate cavin1-cavin2 and cavin1-cavin3 subcomplexes. Each of these subcomplexes contain 9 ± 2 cavin molecules and appear to be the building blocks of the caveolar coat. High resolution immunoelectron microscopy suggests a remarkable nanoscale organization of these separate subcomplexes, forming individual striations on the surface of caveolae. DOI: http://dx.doi.org/10.7554/eLife.01434.001
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spelling pubmed-39031332014-01-29 Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae Gambin, Yann Ariotti, Nicholas McMahon, Kerrie-Ann Bastiani, Michele Sierecki, Emma Kovtun, Oleksiy Polinkovsky, Mark E Magenau, Astrid Jung, WooRam Okano, Satomi Zhou, Yong Leneva, Natalya Mureev, Sergey Johnston, Wayne Gaus, Katharina Hancock, John F Collins, Brett M Alexandrov, Kirill Parton, Robert G eLife Biophysics and Structural Biology In mammalian cells three closely related cavin proteins cooperate with the scaffolding protein caveolin to form membrane invaginations known as caveolae. Here we have developed a novel single-molecule fluorescence approach to directly observe interactions and stoichiometries in protein complexes from cell extracts and from in vitro synthesized components. We show that up to 50 cavins associate on a caveola. However, rather than forming a single coat complex containing the three cavin family members, single-molecule analysis reveals an exquisite specificity of interactions between cavin1, cavin2 and cavin3. Changes in membrane tension can flatten the caveolae, causing the release of the cavin coat and its disassembly into separate cavin1-cavin2 and cavin1-cavin3 subcomplexes. Each of these subcomplexes contain 9 ± 2 cavin molecules and appear to be the building blocks of the caveolar coat. High resolution immunoelectron microscopy suggests a remarkable nanoscale organization of these separate subcomplexes, forming individual striations on the surface of caveolae. DOI: http://dx.doi.org/10.7554/eLife.01434.001 eLife Sciences Publications, Ltd 2014-01-28 /pmc/articles/PMC3903133/ /pubmed/24473072 http://dx.doi.org/10.7554/eLife.01434 Text en Copyright © 2013, Gambin et al http://creativecommons.org/licenses/by/3.0/ This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Biophysics and Structural Biology
Gambin, Yann
Ariotti, Nicholas
McMahon, Kerrie-Ann
Bastiani, Michele
Sierecki, Emma
Kovtun, Oleksiy
Polinkovsky, Mark E
Magenau, Astrid
Jung, WooRam
Okano, Satomi
Zhou, Yong
Leneva, Natalya
Mureev, Sergey
Johnston, Wayne
Gaus, Katharina
Hancock, John F
Collins, Brett M
Alexandrov, Kirill
Parton, Robert G
Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
title Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
title_full Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
title_fullStr Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
title_full_unstemmed Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
title_short Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
title_sort single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae
topic Biophysics and Structural Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3903133/
https://www.ncbi.nlm.nih.gov/pubmed/24473072
http://dx.doi.org/10.7554/eLife.01434
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