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Singlet oxygen photosensitisation by the fluorescent protein Pp2FbFP L30M, a novel derivative of Pseudomonas putida flavin-binding Pp2FbFP

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Abstract

Flavin-binding fluorescent proteins (FbFPs) are a class of fluorescent reporters that have been increasingly used as reporters in the study of cellular structures and dynamics. Flavin’s intrinsic high singlet oxygen (1O2) quantum yield (ΦΔ = 0.51) provides a basis for the development of new FbFP mutants capable of photosensitising 1O2 for mechanistic and therapeutic applications, as recently exemplified by the FbFP miniSOG. In the present work we report an investigation on the 1O2 photoproduction by Pp2FbFP L30M, a novel derivative of Pseudomonas putida Pp2FbFP. Direct detection of 1O2 through its phosphorescence at 1275 nm yielded the value ΦΔ = 0.09 ± 0.01, which is the highest 1O2 quantum yield reported to date for any FP and is approximately 3-fold higher than the ΦΔ for miniSOG. Unlike miniSOG, transient absorption measurements revealed the existence of two independent triplet states each with a different ability to sensitise 1O2.

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Authors and Affiliations

  1. Institut Químic de Sarrià, Universitat Ramon Llull, Via Augusta 390, 08017, Barcelona, Spain

    Joaquim Torra, Andrés Burgos-Caminal, Rubén Ruiz-González & Santi Nonell

  2. Institute of Molecular Enzyme Technology, Heinrich-Heine-University Düsseldorf, Forschungszentrum Jülich, 52425, Jülich, Germany

    Stephan Endres, Marcus Wingen & Thomas Drepper

  3. Institute of Complex Systems 4 (ICS-4, Cellular Biophysics), Forschungszentrum Jülich, 52425, Jülich, Germany

    Thomas Gensch

Authors
  1. Joaquim Torra
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  2. Andrés Burgos-Caminal
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  3. Stephan Endres
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  5. Thomas Drepper
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  6. Thomas Gensch
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  7. Rubén Ruiz-González
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  8. Santi Nonell
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Correspondence to Santi Nonell.

Additional information

Dedicated to Professor Kristian Berg on the occasion of his 60th birthday.

Electronic supplementary information (ESI) available: Pp2FbFP L30M 1O2 phosphorescence kinetics; effect of oxygen concentration on the rate of Pp2FbFP L30M triplet state decay; transient absorbance decays; deconvolution of 1O2 phosphorescence. See DOI: 10.1039/c4pp00338a

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Torra, J., Burgos-Caminal, A., Endres, S. et al. Singlet oxygen photosensitisation by the fluorescent protein Pp2FbFP L30M, a novel derivative of Pseudomonas putida flavin-binding Pp2FbFP. Photochem Photobiol Sci 14, 280–287 (2015). https://doi.org/10.1039/c4pp00338a

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  • DOI: https://doi.org/10.1039/c4pp00338a

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