Photoswitchable Fluorescent Proteins: Mechanisms on Ultrafast Timescales
Photoswitchable Fluorescent Proteins: Mechanisms on Ultrafast Timescales
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DOI：
10.3390/ijms23126459
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发表时间：
2022-06-09
影响因子：
5.6
通讯作者：
Fang, Chong
中科院分区：
文献类型：
--
作者：
Tang, Longteng;Fang, Chong
关键词：
超分辨率成像（SRI）的进展依赖于具有新颖光致变色特性的荧光蛋白。利用光，可逆切换的荧光蛋白（RSFP）可以在亮和暗状态之间转换，用于许多光循环，它们的出现激发了先进SRI技术的发明。一般的光开关机制涉及发色团顺反异构化和质子转移的负和正RSFP和水合脱水的去耦RSFP。然而，对这些过程在超快时间尺度（飞秒到毫秒）上的详细了解还缺乏，这从根本上阻碍了RSFP的进一步发展。本文综述了近年来利用超快电子光谱、振动光谱和时间分辨晶体学研究RSFPs光开关的研究进展。我们发现，一些研究充分的蛋白质已经获得了显着的见解，但实时的“行动”的双向顺反异构化，质子转移，和中间状态的细节仍然不清楚，大多数系统，和许多其他相关的蛋白质尚未研究。我们希望这篇综述能为现有和未来的工程RSFP奠定基础，并激发更多的超快研究。所获得的机理见解将加速RSFP的合理开发，具有增强的双向开关速率和效率，更好的光稳定性，更高的亮度和更红的发射颜色。
The advancement of super-resolution imaging (SRI) relies on fluorescent proteins with novel photochromic properties. Using light, the reversibly switchable fluorescent proteins (RSFPs) can be converted between bright and dark states for many photocycles and their emergence has inspired the invention of advanced SRI techniques. The general photoswitching mechanism involves the chromophore cis-trans isomerization and proton transfer for negative and positive RSFPs and hydration–dehydration for decoupled RSFPs. However, a detailed understanding of these processes on ultrafast timescales (femtosecond to millisecond) is lacking, which fundamentally hinders the further development of RSFPs. In this review, we summarize the current progress of utilizing various ultrafast electronic and vibrational spectroscopies, and time-resolved crystallography in investigating the on/off photoswitching pathways of RSFPs. We show that significant insights have been gained for some well-studied proteins, but the real-time “action” details regarding the bidirectional cis-trans isomerization, proton transfer, and intermediate states remain unclear for most systems, and many other relevant proteins have not been studied yet. We expect this review to lay the foundation and inspire more ultrafast studies on existing and future engineered RSFPs. The gained mechanistic insights will accelerate the rational development of RSFPs with enhanced two-way switching rate and efficiency, better photostability, higher brightness, and redder emission colors.
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影响因子：
3.5
作者：
Cormack, BP;Valdivia, RH;Falkow, S
通讯作者：
Falkow, S
影响因子：
3.4
作者：
Ando, Ryoko;Flors, Cristina;Miyawaki, Atsushi
通讯作者：
Miyawaki, Atsushi
影响因子：
4
作者：
Arai, Yoshiyuki;Takauchi, Hiroki;Nagai, Takeharu
通讯作者：
Nagai, Takeharu
影响因子：
56.9
作者：
CHALFIE, M;TU, Y;PRASHER, DC
通讯作者：
PRASHER, DC
DOI：
10.1073/pnas.0700629104
发表时间：
2007-08-07
影响因子：
11.1
作者：
Andresen, Martin;Stiel, Andre C.;Jakobs, Stefan
通讯作者：
Jakobs, Stefan