ID | 60986 |
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著者 |
Kim, Hyungjin
Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Watanabe, Sho
Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University
Watanabe, Kazunori
Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University
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Ohtsuki, Takashi
Department of Interdisciplinary Science and Engineering in Health Systems, Okayama University
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抄録 | Investigation of the relevance between cell cycle status and the bioactivity of exogenously delivered biomacromolecules is hindered by their time-consuming cell internalization and the cytotoxicity of transfection methods. In this study, we addressed these problems by utilizing the photochemical internalization (PCI) method using a peptide/protein-photosensitizer conjugate, which enables immediate cytoplasmic internalization of the bioactive peptides/proteins in a light-dependent manner with low cytotoxicity. To identify the cell-cycle dependent apoptosis, a TatBim peptide-photosensitizer conjugate (TatBim-PS) with apoptotic activity was photo-dependently internalized into HeLa cells expressing a fluorescent ubiquitination-based cell cycle indicator (Fucci2). Upon irradiation, cytoplasmic TatBim-PS internalization exceeded 95% for all cells classified in the G(1), S, and G(2)/M cell cycle phases with no significant differences between groups. TatBim-PS-mediated apoptosis was more efficiently triggered by photoirradiation in the G(1)/S transition than in the G(1) and S/G(2)/M phases, suggesting high sensitivity of the former phase to Bim-induced apoptosis. Thus, the cell cycle dependence of Bim peptide-induced apoptosis was successfully investigated using Fucci2 indicator and the PCI method. Since PCI-mediated cytoplasmic internalization of peptides is rapid and does not span multiple cell cycle phases, the Fucci-PCI method constitutes a promising tool for analyzing the cell cycle dependence of peptides/protein functions.
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キーワード | Biological techniques
Biotechnology
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発行日 | 2020-11-05
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出版物タイトル |
Scientific Reports
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巻 | 10巻
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号 | 1号
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出版者 | Nature Research
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開始ページ | 19087
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ISSN | 2045-2322
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資料タイプ |
学術雑誌論文
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言語 |
英語
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OAI-PMH Set |
岡山大学
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著作権者 | © The Author(s) 2020
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論文のバージョン | publisher
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PubMed ID | |
DOI | |
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関連URL | isVersionOf https://doi.org/10.1038/s41598-020-76100-7
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ライセンス | http://creat iveco mmons .org/licen ses/by/4.0/
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助成機関名 |
日本学術振興会
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助成番号 | JP18H02090
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