谷胱甘肽
化学
活体细胞成像
合理设计
费斯特共振能量转移
荧光
生物物理学
荧光寿命成像显微镜
动力学
细胞内
细胞
纳米技术
生物化学
酶
量子力学
生物
物理
材料科学
作者
Keitaro Umezawa,Masafumi Yoshida,Mako Kamiya,Tatsuya Yamasoba,Yasuteru Urano
出处
期刊:Nature Chemistry
[Springer Nature]
日期:2016-11-07
卷期号:9 (3): 279-286
被引量:427
摘要
Alterations in glutathione (GSH) homeostasis are associated with a variety of diseases and cellular functions, and therefore, real-time live-cell imaging and quantification of GSH dynamics are important for understanding pathophysiological processes. However, existing fluorescent probes are unsuitable for these purposes due to their irreversible fluorogenic mechanisms or slow reaction rates. In this work, we have successfully overcome these problems by establishing a design strategy inspired by Mayr's work on nucleophilic reaction kinetics. The synthesized probes exhibit concentration-dependent, reversible and rapid absorption/fluorescence changes (t1/2 = 620 ms at [GSH] = 1 mM), as well as appropriate Kd values (1-10 mM: within the range of intracellular GSH concentrations). We also developed FRET-based ratiometric probes, and demonstrated that they are useful for quantifying GSH concentration in various cell types and also for real-time live-cell imaging of GSH dynamics with temporal resolution of seconds.
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