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Int. J. Mol. Sci. 2017, 18(6), 1187; doi:10.3390/ijms18061187

Deciphering Structural Photophysics of Fluorescent Proteins by Kinetic Crystallography

Institut de Biologie Structurale, Univ. Grenoble Alpes, CNRS, CEA, CNRS, IBS, F-38000 Grenoble, France
Academic Editor: Kevin D. Belfield
Received: 25 April 2017 / Revised: 24 May 2017 / Accepted: 26 May 2017 / Published: 2 June 2017
(This article belongs to the Special Issue Fluorescent Proteins)
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Abstract

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. View Full-Text
Keywords: fluorescent proteins; phototransformations; protein dynamics; in crystallo optical spectroscopy; kinetic X-ray crystallography; quantum mechanics/molecular mechanics; single molecule imaging fluorescent proteins; phototransformations; protein dynamics; in crystallo optical spectroscopy; kinetic X-ray crystallography; quantum mechanics/molecular mechanics; single molecule imaging
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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Bourgeois, D. Deciphering Structural Photophysics of Fluorescent Proteins by Kinetic Crystallography. Int. J. Mol. Sci. 2017, 18, 1187.

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