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Photophysics and photochemistry of dyes bound to human serum albumin are determined by the dye localization

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Abstract

The photophysics and photochemistry of rose bengal (RB) and methylene blue (MB) bound to human serum albumin (HSA) have been investigated under a variety of experimental conditions. Distribution of the dyes between the external solvent and the protein has been estimated by physical separation and fluorescence measurements. The main localization of protein-bound dye molecules was estimated by the intrinsic fluorescence quenching, displacement of fluorescent probes bound to specific protein sites, and by docking modelling. All the data indicate that, at low occupation numbers, RB binds strongly to the HSA site I, while MB localizes predominantly in the protein binding site II. This different localization explains the observed differences in the dyes’ photochemical behaviour. In particular, the environment provided by site I is less polar and considerably less accessible to oxygen. The localization of RB in site I also leads to an efficient quenching of the intrinsic protein fluorescence (ascribed to the nearby Trp residue) and the generation of intra-protein singlet oxygen, whose behaviour is different to that observed in the external solvent or when it is generated by bound MB.

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Authors and Affiliations

  1. Facultad de Química, Departamento de Química Física, Pontificia Universidad Católica de Chile, Santiago, Chile

    Emilio Alarcón & Ana Maria Edwards

  2. Departamento de Ciencias del Ambiente, Facultad de Química y Biología, Universidad de Santiago de Chile, USACH, Av. Bernardo O’Higgins, 3363, Santiago, Chile

    Alexis Aspee & Eduardo A. Lissi

  3. Instituto de Química del NOA (INQUINOA-CONICET), Facultad de Agronomía y Agroindustrias, Universidad Nacional de Santiago del Estero, Santiago del Estero, Argentina

    Faustino E. Moran & Claudio D. Borsarelli

  4. Centro de Bio-informática, Universidad de Talca, Chile

    Danilo Gonzalez-Nilo & Horacio Poblete

  5. Faculty of Science, University of Ottawa, 10, Marie Curie, K1N 6N5, Canada

    J. C. Scaiano

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  1. Emilio Alarcón
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  2. Ana Maria Edwards
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Correspondence to Eduardo A. Lissi.

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Alarcón, E., Edwards, A.M., Aspee, A. et al. Photophysics and photochemistry of dyes bound to human serum albumin are determined by the dye localization. Photochem Photobiol Sci 9, 93–102 (2010). https://doi.org/10.1039/b9pp00091g

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