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Semi-retentive cytoskeletal fractionation (SERCYF): A novel method for the biochemical analysis of the organization of microtubule and actin cytoskeleton networks

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Title: Semi-retentive cytoskeletal fractionation (SERCYF): A novel method for the biochemical analysis of the organization of microtubule and actin cytoskeleton networks
Authors: Sato, Yuta Browse this author
Murakami, Yota Browse this author
Takahashi, Masayuki Browse this author →KAKEN DB
Keywords: Cytoskeleton
Nonmuscle myosin II
Issue Date: 8-Jul-2017
Publisher: Elsevier
Journal Title: Biochemical and biophysical research communications
Volume: 488
Issue: 4
Start Page: 614
End Page: 620
Publisher DOI: 10.1016/j.bbrc.2017.05.083
PMID: 28526408
Abstract: A variety of biochemical fractionation methods are available for the quantification of cytoskeletal components. However, each method is designed to target only one cytoskeletal network, either the micro tubule (MT) or actin cytoskeleton, and non-targeted cytoskeletal networks are ignored. Considering the importance of MT actin crosstalk, the organization of both the targeted and non-targeted cytoskeletal networks must be retained intact during fractionation for the accurate analysis of cytoskeletal organization. In this study, we reveal that existing fractionation methods, represented by the MT sedimentation-method for MTs and the Triton X-100 solubility assay-method for actin cytoskeletons, disrupt the organizations of the non-targeted cytoskeletons. We demonstrate a novel fractionation method for the accurate analysis of the cytoskeletal organizations using a taxol-containing PEM-based permeabilization buffer, which we name "semi-retentive cytoskeletal fractionation (SERCYF)-method". The organizations of both MTs and actin cytoskeletons were retained intact even after permeabilization with this buffer. By using the SERCYF-method, we analyzed the effects of nocodazole on the cytoskeletal organizations biochemically and showed promotion of the actin cytoskeletal organization by MT depolymerization. (C) 2017 Elsevier Inc. All rights reserved.
Rights: © 2017 This manuscript version is made available under the CC-BY-NC-ND 4.0 license
Type: article (author version)
Appears in Collections:理学院・理学研究院 (Graduate School of Science / Faculty of Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 高橋 正行

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