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Excited state dynamics of the photoconvertible fluorescent protein Kaede revealed by ultrafast spectroscopy

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Abstract

The ultrafast excited state dynamics of the fluorescent protein Kaede has been investigated by employing time resolved fluorescence and transient absorption. Upon irradiation of its neutral state, the protein undergoes an efficient conversion to a state that fluoresces at longer wavelengths. The molecular basis of the photoconversion involves an expansion of the chromophore π-conjugation by formal β-elimination but details of the reaction pathway remain subject to debate. Based on the kinetics observed in experiments on the protein sample in both H2O and D2O buffers, we suggest that a light-initiated cleavage mechanism (20 ps) could take place, forming the neutral red state in which the red chromophore resides. Excitation of the neutral red form results in the formation of the red anionic species via two Förster resonance energy transfer (FRET) channels. FRET between red neutral and red anionic forms occurs within the tetramer with time constants of 13.4 ps and 210 ps. In contrast to literature proposals no ESPT was observed.

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Authors and Affiliations

  1. Laboratory of Photochemistry and Spectroscopy, Department of Chemistry, KU Leuven, Celestijnenlaan 200F, 3001, Heverlee, Belgium

    Eduard Fron, Susana Rocha, Peter Dedecker & Johan Hofkens

  2. Laboratoire de Spectrochimie Infrarouge et Raman, Université Lille Nord de France, Lille 1, LASIR, F-59655 Villeneuve d’Ascq, CNRS-UMR, 8516, France

    Michel Sliwa

  3. Institut de Biologie Structurale (IBS), Université Grenoble Alpes, F-38027, Grenoble, France

    Virgile Adam

  4. CNRS, IBS, F-38027, Grenoble, France

    Virgile Adam

  5. CEA, DSV, IBS, F-38027, Grenoble, France

    Virgile Adam

  6. Centre of Microbial and Plant Genetics, Faculty of Bioscience Engineering, KU Leuven, Kasteelpark Arenberg 20, 3001, Heverlee, Belgium

    Jan Michiels

  7. Laboratory of Biomolecular Network Dynamics, Department of Chemistry, KU Leuven, Celestijnenlaan 200G, 3001, Heverlee, Belgium

    Hideaki Mizuno

Authors
  1. Eduard Fron
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  2. Michel Sliwa
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  3. Virgile Adam
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  4. Jan Michiels
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  5. Susana Rocha
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  6. Peter Dedecker
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  7. Johan Hofkens
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  8. Hideaki Mizuno
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Corresponding author

Correspondence to Hideaki Mizuno.

Additional information

Electronic supplementary information (ESI) available: Experimental details and additional results are presented. See DOI: 10.1039/c3pp50335f

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Fron, E., Sliwa, M., Adam, V. et al. Excited state dynamics of the photoconvertible fluorescent protein Kaede revealed by ultrafast spectroscopy. Photochem Photobiol Sci 13, 867–874 (2014). https://doi.org/10.1039/c3pp50335f

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  • DOI: https://doi.org/10.1039/c3pp50335f

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