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Real-time monitoring of the in vivo redox state transition using the ratiometric redox state sensor protein FROG/B.
Proceedings of the National Academy of Sciences of the United States of America ( IF 11.1 ) Pub Date : 2020-07-07 , DOI: 10.1073/pnas.1918919117
Kazunori Sugiura 1 , Shoko Mihara 1, 2 , Nae Fu 1, 2 , Toru Hisabori 2, 3
Affiliation  

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



中文翻译:

使用比例氧化还原状态传感器蛋白FROG / B实时监测体内氧化还原状态转变。

细胞内氧化还原状态是​​调节细胞中各种生理现象的关键因素之一。因此,监视此状态对于了解细胞中的生理稳态非常重要。已经开发了各种荧光传感器蛋白来监测细胞内氧化还原状态。我们还开发了名为Oba-Q和Re-Q的荧光氧化还原传感器蛋白,它们的发射分别在氧化和还原条件下淬灭。尽管这些传感器可用于可视化随时间变化的细胞氧化还原变化,但它们的缺点是其发射信号直接受其原位表达水平的影响。为了克服这个问题,我们开发了具有单个激发峰和双重可变发射峰的氧化还原传感器蛋白。该传感器蛋白在氧化条件下显示绿色发射,在还原条件下显示蓝色发射。因此,我们将这种传感器命名为FROG / B,荧光蛋白,其氧化还原依赖性绿色/蓝色变化。通过使用该传感器,我们仅通过计算绿色和蓝色信号之间的发射比率就成功地定量测量了由明/暗过渡引起的蓝细菌细胞内细胞内氧化还原电位的变化。

更新日期:2020-07-08
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