Photoactivatable Fluorescent Protein mEos2 Displays Repeated Photoactivation after a Long-Lived Dark State in the Red Photoconverted Form
Photoactivatable Fluorescent Protein mEos2 Displays Repeated Photoactivation after a Long-Lived Dark State in the Red Photoconverted Form
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DOI：
10.1021/jz1003523
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发表时间：
2010-05-06
影响因子：
5.7
通讯作者：
Radenovic, A.
中科院分区：
文献类型：
--
作者：
Annibale, P.;Scarselli, M.;Radenovic, A.
使用 405 nm 光照明可以恢复已进入长寿命黑暗状态的单个绿色荧光蛋白 (GFP) 突变体的发射。所报告的行为是反向光开关蛋白 Dronpa 及其突变体的标准。然而，关于 mEos2 光激活荧光蛋白 (PA-FP) 的传统知识是，一旦漂白，这种荧光团就很难重新激活，除了少数群体可能表现出这种行为。在这里，我们表明，在典型的实验中，大约 50% 的所研究的单分子时间轨迹显示出多次重新激活，这使得这成为 mEos2 PA-FP 看似固有的特征。这些结果暗示了 mEos2 和其他可逆光活化探针（例如 Dronpa）之间的一些相似之处。我们在光激活定位显微镜（PALM）实验的框架中研究了这种现象的后果。
Illumination with 405 nm light can recover the emission for single green fluorescent protein (GFP) mutants that have gone into a long-lived dark state. The reported behavior is the standard for the reverse photoswitchable protein Dronpa and its mutants. However, conventional knowledge regarding the mEos2 photoactivatable fluorescent protein (PA-FP) is that, once bleached, this fluorophore is hardly reactivated, aside from a minority population that might display this behavior. Here we show that, in a typical experiment approximately 50% of the investigated single molecule time traces display multiple reactivations, making this a seemingly inherent feature of the mEos2 PA-FP. These results hint to some similarities between mEos2 and other reversibly photoactivatable probes such as Dronpa. We investigate the consequences of this phenomenon in the framework of photoactivated localization microscopy (PALM) experiments.