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PNAS:细胞内氧化还原状态实时监控的感应蛋白构建

已有 525 次阅读 2020-7-12 20:02 |个人分类:每日摘要|系统分类:论文交流

Real-time monitoring of the in vivo redox state transition using the ratiometric redox state sensor protein FROG/B

第一作者Kazunori Sugiura

第一单位东京工业大学

通讯作者Toru Hisabori


 Abstract 


背景回顾The intracellular redox state is one of the key factors regulating various physiological phenomena in the cell. Monitoring this state is therefore important for understanding physiological homeostasis in cells. Various fluorescent sensor proteins have already been developed to monitor intracellular redox state. We also developed fluorescent redox sensor proteins named Oba-Q and Re-Q, the emissions of which are quenched under oxidized and reduced conditions, respectively.


提出问题:Although these sensors were useful to visualize the redox changes in the cell over time, they have the weakness that their emission signals are directly influenced by their in situ expression levels.


解决问题:To overcome this problem, we developed a redox sensor protein with a single excitation peak and dual variable emission peaks.


机制:This sensor protein shows green emission under oxidized conditions and blue emission under reduced conditions. We therefore named this sensor FROG/B, fluorescent protein with redox-dependent change in green/blue.


实例:By using this sensor, we successfully measured the changes in intracellular redox potentials in cyanobacterial cells quantitatively caused by light/dark transition just by calculating the ratio of emission between green and blue signals.


 摘  要 


细胞内的氧化还原状态是调控多个生理现象的关键因素。因此,监控细胞内的氧化还原状态对于理解细胞的生理内稳态至关重要。目前已经有研究报道了多个荧光感应蛋白来监控细胞内的氧化还原状态。作者也开发了荧光感应蛋白,叫做Oba-QRe-Q,其发射光会分别在氧化和还原条件下淬灭。尽管这些感应器有助于可视化细胞内的氧化还原状态随时间的变化动态,但仍旧有缺点,其发射信号直接受到原位表达水平的影响。为了克服这个缺陷,作者又开发了一个氧化还原感应蛋白,其具有单个激发峰和两个不同的发射峰。该感应蛋白会在氧化条件下发射绿光,而在还原条件下发射蓝光。因此,作者将该感应蛋白命名为FROG/B,即依赖于氧化还原状态的绿色/蓝色荧光蛋白。利用该感应蛋白,作者通过计算绿色和蓝色信号之间的发射比率成功测量了明-暗过渡所引起的蓝细菌胞内氧化还原电势的变化。


 通讯作者 


**Toru Hisabori**


个人简介:

1982年,早稻田大学,硕士

1985年,早稻田大学,博士


研究方向:

叶绿体ATP合酶的调节和光合功能的氧化还原控制机制


doi: 10.1073/pnas.1918919117


Journal: PNAS

Published date: July 07, 2020



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