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Title: Real-time TIRF Observation of Vinculin Reqruitment to Stretched α-catenin by AFM
Authors: Maki, Koichiro  kyouindb  KAKEN_id
Han, Sung-Woong
Hirano, Yoshinori
Yonemura, Shigenobu
Hakoshima, Toshio
Adachi, Taiji  kyouindb  KAKEN_id  orcid https://orcid.org/0000-0001-5280-4156 (unconfirmed)
Author's alias: 牧, 功一郎
安達, 泰治
Issue Date: 25-Jan-2018
Publisher: Springer Nature
Journal title: Scientific Reports
Volume: 8
Thesis number: 1575
Abstract: 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, α-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 α-catenin-mediated mechanotransduction is a dynamic molecular process, which involves a conformational change of α-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, α-catenin molecules (residues 276–634; the mechano-sensitive M₁-M₃ 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 α-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.
Rights: © The Author(s) 2018. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
URI: http://hdl.handle.net/2433/234504
DOI(Published Version): 10.1038/s41598-018-20115-8
PubMed ID: 29371682
Appears in Collections:Journal Articles

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