Issue 2, 2010
From the journal:

Photochemical & Photobiological Sciences

Low-temperature switching by photoinduced protonation in photochromic fluorescent proteins†

Aline Regis Faro,a   Virgile Adam,b   Philippe Carpentier,a   Claudine Darnault,a   Dominique Bourgeois*a  and  Eve de Rosny*a  

* Corresponding authors

a IBS, Institut de Biologie Structurale Jean-Pierre Ebel, CEA, CNRS, Université Joseph Fourier, 41 rue Jules Horowitz, Grenoble, France
E-mail: dominique.bourgeois@ibs.fr, eve.derosny@ibs.fr
Fax: +33-4-38785122
Tel: +33-4-38789644

b ESRF, 6 rue Jules Horowitz, BP 220, Grenoble Cedex, France

Abstract

We have studied the photoswitching behaviour of a number of photochromic fluorescent proteins at cryo-temperature. Spectroscopic investigations at the ensemble level showed that EYFP, Dronpa and IrisFP all exhibit reversible photoswitching at 100 K, albeit with a low quantum yield. The photophysics of the process were studied in more details in the case of EYFP. The data suggest that photoinduced protonation of the chromophore is responsible for off-switching at cryo-temperature, and thus is possible in the absence of significant conformational freedom. This finding is consistent with the hypothesis that chromophore protonation may precede large amplitude conformational changes such as cis–trans isomerisation during off-photoswitching at room temperature. However, our data suggest that low-barrier photoinduced protonation pathways may in fact compete with room-temperature off-switching reactions in photochromic fluorescent proteins. The occurrence of reversible photoswitching at low-temperature is of interest to envisage cryo-nanoscopy experiments using genetically encoded fluorophores.

Graphical abstract: Low-temperature switching by photoinduced protonation in photochromic fluorescent proteins

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Article information

DOI
https://doi.org/10.1039/B9PP00121B
Article type
Paper
Submitted
02 Oct 2009
Accepted
23 Nov 2009
First published
19 Jan 2010

Photochem. Photobiol. Sci., 2010,9, 254-262

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Low-temperature switching by photoinduced protonation in photochromic fluorescent proteins

A. R. Faro, V. Adam, P. Carpentier, C. Darnault, D. Bourgeois and E. de Rosny, Photochem. Photobiol. Sci., 2010, 9, 254 DOI: 10.1039/B9PP00121B

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