title: Photoinduced Chromophore Hydration in the Fluorescent Protein Dreiklang Is Triggered by Ultrafast Excited-State Proton Transfer Coupled to a Low-Frequency Vibration.
abstract: Because of growing applications in advanced fluorescence imaging, the mechanisms and dynamics of photoinduced reactions in reversibly photoswitchable fluorescent proteins are currently attracting much interest. We report the first time-resolved study of the photoswitching of Dreiklang, so far the only fluorescent protein to undergo reversible photoinduced chromophore hydration. Using broadband femtosecond transient absorption spectroscopy, we show that the reaction is triggered by an ultrafast deprotonation of the chromophore phenol group in the excited state in 100 fs. This primary step is accompanied by coherent oscillations that we assign to its coupling with a low-frequency mode, possibly a deformation of the chromophore hydrogen bond network. A ground-state intermediate is formed in the picosecond-nanosecond regime that we tentatively assign to the deprotonated water adduct. We suggest that proton ejection from the phenol group leads to a charge transfer from the phenol to the imidazolinone ring, which triggers imidazolinone protonation by nearby Glu222 and catalyzes the addition of the water molecule.
authors.affiliation: Ecole normale supérieure, PSL Research University , Département de Chimie, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|CNRS , PASTEUR, 24 rue Lhomond, 75005 Paris, France.
authors.forename: Fabien
authors.initials: F
authors.lastname: Lacombat
authors.affiliation: Ecole normale supérieure, PSL Research University , Département de Chimie, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|CNRS , PASTEUR, 24 rue Lhomond, 75005 Paris, France.
authors.forename: Pascal
authors.identifier: http://orcid.org/0000-0002-8021-4521
authors.initials: P
authors.lastname: Plaza
authors.affiliation: Ecole normale supérieure, PSL Research University , Département de Chimie, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|CNRS , PASTEUR, 24 rue Lhomond, 75005 Paris, France.
authors.forename: Marie-Aude
authors.initials: MA
authors.lastname: Plamont
authors.affiliation: Ecole normale supérieure, PSL Research University , Département de Chimie, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|Sorbonne Universités, UPMC Univ Paris 06, PASTEUR, 24 rue Lhomond, 75005 Paris, France.|CNRS , PASTEUR, 24 rue Lhomond, 75005 Paris, France.
authors.forename: Agathe
authors.identifier: http://orcid.org/0000-0002-2333-0227
authors.initials: A
authors.lastname: Espagne