Bourgeois Dominique
Institut de Biologie Structurale, Univ. Grenoble Alpes, CNRS, CEA, CNRS, IBS, F-38000 Grenoble, France.
Int J Mol Sci. 2017 Jun 2;18(6):1187. doi: 10.3390/ijms18061187.
Because they enable labeling of biological samples in a genetically-encoded manner, Fluorescent Proteins (FPs) have revolutionized life sciences. Photo-transformable fluorescent proteins (PTFPs), in particular, recently attracted wide interest, as their fluorescence state can be actively modulated by light, a property central to the emergence of super-resolution microscopy. PTFPs, however, exhibit highly complex photophysical behaviours that are still poorly understood, hampering the rational engineering of variants with improved performances. We show that kinetic crystallography combined with in crystallo optical spectroscopy, modeling approaches and single-molecule measurements constitutes a powerful tool to decipher processes such as photoactivation, photoconversion, photoswitching, photoblinking and photobleaching. Besides potential applications for the design of enhanced PTFPs, these investigations provide fundamental insight into photoactivated protein dynamics.
由于荧光蛋白（FPs）能够以基因编码的方式对生物样品进行标记，因此彻底改变了生命科学。特别是可光转换荧光蛋白（PTFPs），最近引起了广泛关注，因为它们的荧光状态可以通过光进行主动调节，这是超分辨率显微镜出现的核心特性。然而，PTFPs表现出高度复杂的光物理行为，目前仍知之甚少，这阻碍了具有改进性能的变体的合理工程设计。我们表明，动力学晶体学与晶体光学光谱、建模方法和单分子测量相结合，构成了一个强大的工具，用于破译诸如光激活、光转换、光开关、光闪烁和光漂白等过程。除了在设计增强型PTFPs方面的潜在应用外，这些研究还为光激活蛋白动力学提供了基本的见解。
