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Force transduction by Triton cytoskeletons

Force-initiated signal transduction can occur either via membrane-based ionic mechanisms or through changes in cytoskeletal–matrix linkages. We report here the stretch-dependent binding of cytoplasmic proteins to Triton X-100 cytoskeletons of L-929 cells grown on collagen-coated silicone. Triton X-1...

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Detalles Bibliográficos
Autores principales: Sawada, Yasuhiro, Sheetz, Michael P.
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 2002
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2174068/
https://www.ncbi.nlm.nih.gov/pubmed/11839769
http://dx.doi.org/10.1083/jcb.200110068
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author Sawada, Yasuhiro
Sheetz, Michael P.
author_facet Sawada, Yasuhiro
Sheetz, Michael P.
author_sort Sawada, Yasuhiro
collection PubMed
description Force-initiated signal transduction can occur either via membrane-based ionic mechanisms or through changes in cytoskeletal–matrix linkages. We report here the stretch-dependent binding of cytoplasmic proteins to Triton X-100 cytoskeletons of L-929 cells grown on collagen-coated silicone. Triton X-100–insoluble cytoskeletons were stretched by 10% and incubated with biotinylated cytoplasmic proteins. Analysis with two-dimensional gel electrophoresis showed stretch-dependent binding of more than 10 cytoplasmic protein spots. Bound cytoplasmic proteins were purified by a photocleavable biotin tag and stretch-dependent binding of paxillin, focal adhesion kinase, and p130Cas was found, whereas the binding of vinculin was unchanged and actin binding decreased with stretch. Paxillin binding upon stretch was morphologically and biochemically similar in vitro and in vivo, that is, enhanced in the periphery and inhibited by the tyrosine phosphatase inhibitor, phenylarsine oxide. Thus, we suggest that transduction of matrix forces occurs through force-dependent conformation changes in the integrated cytoskeleton.
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spelling pubmed-21740682008-05-01 Force transduction by Triton cytoskeletons Sawada, Yasuhiro Sheetz, Michael P. J Cell Biol Report Force-initiated signal transduction can occur either via membrane-based ionic mechanisms or through changes in cytoskeletal–matrix linkages. We report here the stretch-dependent binding of cytoplasmic proteins to Triton X-100 cytoskeletons of L-929 cells grown on collagen-coated silicone. Triton X-100–insoluble cytoskeletons were stretched by 10% and incubated with biotinylated cytoplasmic proteins. Analysis with two-dimensional gel electrophoresis showed stretch-dependent binding of more than 10 cytoplasmic protein spots. Bound cytoplasmic proteins were purified by a photocleavable biotin tag and stretch-dependent binding of paxillin, focal adhesion kinase, and p130Cas was found, whereas the binding of vinculin was unchanged and actin binding decreased with stretch. Paxillin binding upon stretch was morphologically and biochemically similar in vitro and in vivo, that is, enhanced in the periphery and inhibited by the tyrosine phosphatase inhibitor, phenylarsine oxide. Thus, we suggest that transduction of matrix forces occurs through force-dependent conformation changes in the integrated cytoskeleton. The Rockefeller University Press 2002-02-18 /pmc/articles/PMC2174068/ /pubmed/11839769 http://dx.doi.org/10.1083/jcb.200110068 Text en Copyright © 2002, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Report
Sawada, Yasuhiro
Sheetz, Michael P.
Force transduction by Triton cytoskeletons
title Force transduction by Triton cytoskeletons
title_full Force transduction by Triton cytoskeletons
title_fullStr Force transduction by Triton cytoskeletons
title_full_unstemmed Force transduction by Triton cytoskeletons
title_short Force transduction by Triton cytoskeletons
title_sort force transduction by triton cytoskeletons
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2174068/
https://www.ncbi.nlm.nih.gov/pubmed/11839769
http://dx.doi.org/10.1083/jcb.200110068
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