Photoswitchable Fluorescent Proteins: Do Not Always Look on the Bright Side
Photoswitchable Fluorescent Proteins: Do Not Always Look on the Bright Side
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DOI：
10.1021/acsnano.6b06298
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发表时间：
2016-10-01
期刊：
影响因子：
17.1
通讯作者：
Nienhaus, Gerd Ulrich
中科院分区：
文献类型：
--
作者：
Nienhaus, Karin;Nienhaus, Gerd Ulrich
光活化荧光蛋白（FPs）已成为活体标本纳米观察的重要标记物。在这一期的ACS Nano中，Wang等人提出了一种可逆的光电开关FP， GMars-Q，他们将其推广为分辨率成像的先进标记物，因为它在关闭状态下的残余强度低，并且开关疲劳低。在这里，我们通过一种机制解释了观察到的GMars-Q的特殊光漂白行为，该机制涉及到蛋白质在暗状态下的有效搁置，从而导致低开关疲劳和低残余关闭强度。对具有针对特定成像技术优化特性的新型FP标记物的持续需求。对这类蛋白质进行工程设计的努力将大大受益于对其光物理和光化学机理的深入了解。
Photoactivatable fluorescent proteins (FPs) have become essential markers for nanoscopy on live specimens. In this issue of ACS Nano, Wang et al. present a reversibly photoswitching FP, GMars-Q which they promote as an advanced marker for RESOLFT imaging because of its low residual intensity in the off state and low switching fatigue. Here, we explain the observed peculiar photobleaching behavior of GMars-Q by a mechanism that involves efficient shelving of proteins in dark states, resulting in low switching fatigue and low residual off intensity. There is a continuing demand for novel FP markers with properties optimized for specific imaging techniques. Endeavors to engineer such proteins can greatly benefit from increased efforts to acquire deeper mechanistic understanding of their photophysics and photochemistry.