Watching conformational- and photo-dynamics of single fluorescent proteins in solution.
Watching conformational- and photo-dynamics of single fluorescent proteins in solution.
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DOI：
10.1038/nchem.545
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发表时间：
2010-03
期刊：
影响因子：
21.8
通讯作者：
Moerner, W. E.
中科院分区：
文献类型：
--
作者：
Goldsmith, Randall H.;Moerner, W. E.
在不通过固定化显著改变分子的情况下，很难在长时间内观察单个生物分子的动力学。然而，它可以使用反布朗电动力学（ABEL）陷阱，它允许通过实时电动反馈扩展研究液相生物分子-而无需固定-来完成。本文应用ABEL陷阱对一种重要的光合天线蛋白别藻蓝蛋白（APC）进行了研究。该技术允许观察溶液相APC的单个分子超过一秒。我们观察到一个复杂的荧光强度和寿命之间的关系，不能用简单的静态动力学模型解释。光诱导的构象变化的发生和证据是获得的自发发射寿命，这是通常假设为常数的波动。我们的方法提供了一个新的窗口，荧光蛋白的动态和观察是相关的解释在体内单分子成像实验，细菌光合调节，和生物材料的太阳能收集。
Observing the dynamics of single biomolecules over prolonged time periods is difficult to achieve without significantly altering the molecule through immobilization. It can, however, be accomplished using the Anti-Brownian ELectrokinetic (ABEL) Trap, which allows extended investigation of solution-phase biomolecules - without immobilization -through real-time electrokinetic feedback. Here we apply the ABEL trap to study an important photosynthetic antenna protein, Allophycocyanin (APC). The technique allows the observation of single molecules of solution-phase APC for more than one second. We observe a complex relationship between fluorescence intensity and lifetime that cannot be explained by simple static kinetic models. Light-induced conformational changes are shown to occur and evidence is obtained for fluctuations in the spontaneous emission lifetime, which is typically assumed to be constant. Our methods provide a new window into the dynamics of fluorescent proteins and the observations are relevant for the interpretation of in vivo single-molecule imaging experiments, bacterial photosynthetic regulation, and biomaterials for solar energy harvesting.
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影响因子：
64.8
作者：
Lee, Seok-Jin R.;Escobedo-Lozoya, Yasmin;Szatmari, Erzsebet M.;Yasuda, Ryohei
通讯作者：
Yasuda, Ryohei
影响因子：
2.1
作者：
Berglund, AJ;Mabuchi, H
通讯作者：
Mabuchi, H
DOI：
10.1073/pnas.0601210103
发表时间：
2006-05-30
影响因子：
11.1
作者：
Friedel, Miriam;Baumketner, Andrij;Shea, Joan-Emma
通讯作者：
Shea, Joan-Emma
影响因子：
16.8
作者：
Katona, G;Snijder, A;Neutze, R
通讯作者：
Neutze, R
影响因子：
2.9
作者：
GILSON, MK;HONIG, BH
通讯作者：
HONIG, BH