Low-Temperature Chromophore Isomerization Reveals the Photoswitching Mechanism of the Fluorescent Protein Padron
| Autor: | Philippe Carpentier, Delphine Arcizet, Aline Regis Faro, Isabelle Demachy, Guillaume Pompidor, Dominique Bourgeois, Gabriella Jonasson |
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| Přispěvatelé: | Université Joseph Fourier - Grenoble 1 (UJF), Centre National de la Recherche Scientifique (CNRS), Université Paris-Sud - Paris 11 (UP11), Institut Paul Scherrer (IPS), Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), CEA, ANR [ANR-07-BLAN-0107-01], Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA) |
| Rok vydání: | 2011 |
| Předmět: |
STRUCTURAL BASIS
Models Molecular Protein Conformation [SDV]Life Sciences [q-bio] Protonation Molecular Dynamics Simulation Crystallography X-Ray 010402 general chemistry Photochemistry 01 natural sciences Biochemistry Catalysis 03 medical and health sciences Molecular dynamics RAMAN Colloid and Surface Chemistry Protein structure 030304 developmental biology Luminescent Proteins 0303 health sciences PHOTOCYCLE Chemistry Protein dynamics Temperature Stereoisomerism General Chemistry Chromophore Photochemical Processes Fluorescence DRONPA 0104 chemical sciences STATES NANOSCOPY Isomerization |
| Zdroj: | Journal of the American Chemical Society Journal of the American Chemical Society, American Chemical Society, 2011, 133 (41), pp.16362-16365. ⟨10.1021/ja207001y⟩ Journal of the American Chemical Society, 2011, 133 (41), pp.16362-16365. ⟨10.1021/ja207001y⟩ |
| ISSN: | 1520-5126 0002-7863 |
| DOI: | 10.1021/ja207001y |
| Popis: | International audience; Photoactivatable fluorescent proteins are essential players in nanoscopy approaches based on the super-localization of single molecules. The subclass of reversibly photoswitchable fluorescent proteins typically activate through isomerization of the chromophore coupled with a change in its protonation state. However, the interplay between these two events, the details of photoswitching pathways, and the role of protein dynamics remain incompletely understood. Here, by using a combination of structural and spectroscopic approaches, we discovered two fluorescent intermediate states along the on-switching pathway of the fluorescent protein Padron. The first intermediate can be populated at temperatures as low as 100 K and results from a remarkable trans-cis isomerization of the anionic chromophore taking place within a protein matrix essentially deprived of conformational flexibility. This intermediate evolves in the dark at cryotemperatures to a second structurally similar but spectroscopically distinct anionic intermediate. The final fluorescent state, which consists of a mixture of anionic and neutral chromophores in the cis configuration, is only reached above the glass transition temperature, suggesting that chromophore protonation involves solvent interactions mediated by pronounced dynamical breathing of the protein scaffold. The possibility of efficiently and reversibly photoactivating Padron at cryotemperatures will facilitate the development of advanced super-resolution imaging modalities such as cryonanoscopy. |
| Databáze: | OpenAIRE |
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