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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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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2002
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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. |
format | Text |
id | pubmed-2174068 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2002 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT sawadayasuhiro forcetransductionbytritoncytoskeletons AT sheetzmichaelp forcetransductionbytritoncytoskeletons |