2018年1月
Real-time TIRF observation of vinculin recruitment to stretched alpha-catenin by AFM
SCIENTIFIC REPORTS
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- 巻
- 8
- 号
- 1
- 開始ページ
- 1575
- 終了ページ
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1038/s41598-018-20115-8
- 出版者・発行元
- NATURE PUBLISHING GROUP
Adherens junctions (AJs) adaptively change their intensities in response to intercellular tension; therefore, they integrate tension generated by individual cells to drive multicellular dynamics, such as morphogenetic change in embryos. Under intercellular tension, alpha-catenin, which is a component protein of AJs, acts as a mechano-chemical transducer to recruit vinculin to promote actin remodeling. Although in vivo and in vitro studies have suggested that a-catenin-mediated mechanotransduction is a dynamic molecular process, which involves a conformational change of alpha-catenin under tension to expose a cryptic vinculin binding site, there are no suitable experimental methods to directly explore the process. Therefore, in this study, we developed a novel system by combining atomic force microscopy (AFM) and total internal reflection fluorescence (TIRF). In this system, a-catenin molecules (residues 276-634; the mechano-sensitive M-1-M-3 domain), modified on coverslips, were stretched by AFM and their recruitment of Alexa-labeled full-length vinculin molecules, dissolved in solution, were observed simultaneously, in real time, using TIRF. We applied a physiologically possible range of tensions and extensions to alpha-catenin and directly observed its vinculin recruitment. Our new system could be used in the fields of mechanobiology and biophysics to explore functions of proteins under tension by coupling biomechanical and biochemical information.
- リンク情報
- ID情報
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- DOI : 10.1038/s41598-018-20115-8
- ISSN : 2045-2322
- Web of Science ID : WOS:000423422300009