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若林 憲一 所属 京都産業大学 生命科学部 産業生命科学科 職種 教授 |
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| 言語種別 | 英語 |
| 発行・発表の年月 | 2021/09 |
| 形態種別 | 研究論文 |
| 査読 | 査読あり |
| 標題 | Monitoring cellular redox dynamics using newly developed BRET-based redox sensor proteins. |
| 執筆形態 | その他 |
| 掲載誌名 | The Journal of biological chemistry |
| 掲載区分 | 国外 |
| 巻・号・頁 | 297(4),pp.101186-101186 |
| 著者・共著者 | Nae Fu,Kazunori Sugiura,Kumiko Kondo,Shungo Nakamura,Ken-Ichi Wakabayashi,Toru Hisabori |
| 概要 | Reactive oxygen species are key factors that strongly affect the cellular redox state and regulate various physiological and cellular phenomena. To monitor changes in the redox state, we previously developed fluorescent redox sensors named Re-Q, the emissions of which are quenched under reduced conditions. However, such fluorescent probes are unsuitable for use in the cells of photosynthetic organisms because they require photoexcitation that may change intracellular conditions and induce autofluorescence, primarily in chlorophylls. In addition, the presence of various chromophore pigments may interfere with fluorescence-based measurements due to their strong absorbance. To overcome these problems, we adopted the bioluminescence resonance energy transfer (BRET) mechanism for the sensor and developed two BRET-based redox sensors by fusing CFP- or YFP-based Re-Q with the luminescent protein Nluc. We named the resulting redox-sensitive BRET-based indicator probes "ROBINc" and "ROBINy." ROBINc is pH-insensitive, which is especially vital for observation in photosynthetic organisms. By using these sensors, we successfully observed dynamic redox changes caused by an anticancer agent in HeLa cells and light/dark dependent redox changes in the cells of photosynthetic cyanobacterium Synechocystis sp. PCC 6803. Since the newly developed sensors do not require excitation light, they should be especially useful for visualizing intracellular phenomena caused by redox changes in cells containing colored pigments. |
| DOI | 10.1016/j.jbc.2021.101186 |
| PMID | 34517006 |