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An ATP detection system based on the enzyme reaction with biotin protein ligase
http://hdl.handle.net/10228/0002001479
http://hdl.handle.net/10228/000200147988202f83-f7fd-4172-80f1-b928ef112ef5
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
10449875.pdf (1.9 MB)
Download is available from 2026/10/29.
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| アイテムタイプ | 共通アイテムタイプ(1) | |||||||||||||
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| 公開日 | 2025-03-17 | |||||||||||||
| タイトル | ||||||||||||||
| タイトル | An ATP detection system based on the enzyme reaction with biotin protein ligase | |||||||||||||
| 言語 | en | |||||||||||||
| 著者 |
末田, 慎二
× 末田, 慎二
WEKO
958
× 藤井, 聡
WEKO
15863
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| 著作権関連情報 | ||||||||||||||
| 権利情報 | Copyright (c) 2024 Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies. | |||||||||||||
| 言語 | en | |||||||||||||
| 抄録 | ||||||||||||||
| 内容記述タイプ | Abstract | |||||||||||||
| 内容記述 | Adenosine triphosphate (ATP) is the energy currency of all living organisms and can be used as an indicator for cell proliferation and cytotoxicity. In the present work, we have developed a novel ATP detection system by combining the biotinylation reaction from archaeon Sulfolobus tokodaii with fluorescence resonance energy transfer (FRET). In biotinylation from S. tokodaii, an enzyme known as biotin protein ligase (BPL) forms a very stable complex with its product, biotinylated substrate protein (BCCP). Here, BPL and BCCP were fused to the fluorescent proteins Cerulean and Clover, respectively, and ATP detection was accomplished by monitoring the FRET signal between the two fluorescent proteins, since ATP is an essential component for biotinylation and the tight BPL-BCCP complex is formed only after biotinylation. Using this system, we have succeeded in detecting 5 nM of ATP by biotinylation reaction with 50 nM of each fusion protein. Our method has a characteristic that the signal does not decay for at least 2 h after the start of the reaction, unlike in the case of the luminescence-based assay with luciferase commonly used for the ATP detection. Thus, our system allows for ATP detection which is not significantly constrained by measurement timing. | |||||||||||||
| 言語 | en | |||||||||||||
| 書誌情報 |
en : Analytical Biochemistry 巻 696, p. 115698, 発行日 2024-10-29 |
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| 出版社 | ||||||||||||||
| 出版者 | Elsevier | |||||||||||||
| キーワード | ||||||||||||||
| 言語 | en | |||||||||||||
| 主題Scheme | Other | |||||||||||||
| 主題 | ATP detection | |||||||||||||
| キーワード | ||||||||||||||
| 言語 | en | |||||||||||||
| 主題Scheme | Other | |||||||||||||
| 主題 | FRET | |||||||||||||
| キーワード | ||||||||||||||
| 言語 | en | |||||||||||||
| 主題Scheme | Other | |||||||||||||
| 主題 | Biotinylation | |||||||||||||
| キーワード | ||||||||||||||
| 言語 | en | |||||||||||||
| 主題Scheme | Other | |||||||||||||
| 主題 | Fluorescent protein | |||||||||||||
| キーワード | ||||||||||||||
| 言語 | en | |||||||||||||
| 主題Scheme | Other | |||||||||||||
| 主題 | Enzyme reaction | |||||||||||||
| 言語 | ||||||||||||||
| 言語 | eng | |||||||||||||
| 資源タイプ | ||||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||||
| 資源タイプ | journal article | |||||||||||||
| 出版タイプ | ||||||||||||||
| 出版タイプ | AM | |||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_ab4af688f83e57aa | |||||||||||||
| DOI | ||||||||||||||
| 識別子タイプ | DOI | |||||||||||||
| 関連識別子 | https://doi.org/10.1016/j.ab.2024.115698 | |||||||||||||
| ISSN | ||||||||||||||
| 収録物識別子タイプ | PISSN | |||||||||||||
| 収録物識別子 | 0003-2697 | |||||||||||||
| ISSN | ||||||||||||||
| 収録物識別子タイプ | EISSN | |||||||||||||
| 収録物識別子 | 1096-0309 | |||||||||||||
| 査読の有無 | ||||||||||||||
| 値 | yes | |||||||||||||
| 研究者情報 | ||||||||||||||
| URL | https://hyokadb02.jimu.kyutech.ac.jp/html/280_ja.html | |||||||||||||
| 研究者情報 | ||||||||||||||
| URL | https://hyokadb02.jimu.kyutech.ac.jp/html/287_ja.html | |||||||||||||
| 論文ID（連携） | ||||||||||||||
| 値 | 10449875 | |||||||||||||
| 連携ID | ||||||||||||||
| 値 | 14106 | |||||||||||||
