NMR Reveals Light-Induced Changes in the Dynamics of a Photoswitchable Fluorescent Protein.

Authors:
Dr. Virgile Adam, PhD
Dr. Virgile Adam, PhD
CNRS / Institute for Structural Biology (IBS)
Research scientist
single molecule fluorescence microscopy, fluorescent protein engineering, X-ray crystallography
Grenoble | France

Biophys J 2019 Dec 2;117(11):2087-2100. Epub 2019 Nov 2.

Institut de Biologie Structurale, Université Grenoble Alpes, CEA, CNRS, Grenoble, France. Electronic address:

The availability of fluorescent proteins with distinct phototransformation properties is crucial for a wide range of applications in advanced fluorescence microscopy and biotechnology. To rationally design new variants optimized for specific applications, a detailed understanding of the mechanistic features underlying phototransformation is essential. At present, little is known about the conformational dynamics of fluorescent proteins at physiological temperature and how these dynamics contribute to the observed phototransformation properties. Here, we apply high-resolution NMR spectroscopy in solution combined with in situ sample illumination at different wavelengths to investigate the conformational dynamics of rsFolder, a GFP-derived protein that can be reversibly switched between a green fluorescent state and a nonfluorescent state. Our results add a dynamic view to the static structures obtained by x-ray crystallography. Including a custom-tailored NMR toolbox in fluorescent protein research provides new opportunities for investigating the effect of mutations or changes in the environmental conditions on the conformational dynamics of phototransformable fluorescent proteins and their correlation with the observed photochemical and photophysical properties.

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Source
http://dx.doi.org/10.1016/j.bpj.2019.10.035DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6895687PMC
December 2019
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