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

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 fl...

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Published in:Photochemical & photobiological sciences 2010, Vol.9 (1), p.93-102
Main Authors: Alarcón, Emilio, Edwards, Ana Maria, Aspee, Alexis, Moran, Faustino E., Borsarelli, Claudio D., Lissi, Eduardo A., Gonzalez-Nilo, Danilo, Poblete, Horacio, Scaiano, J. C.
Format: Article
Language:English
Subjects:
Binding Sites
Biochemistry
Biomaterials
Chemistry
Coloring Agents - chemistry
Coloring Agents - metabolism
Humans
Methylene Blue - chemistry
Methylene Blue - metabolism
Models, Molecular
Photochemical Processes
Physical Chemistry
Plant Sciences
Protein Conformation
Rose Bengal - chemistry
Rose Bengal - metabolism
Serum Albumin - chemistry
Serum Albumin - metabolism
Singlet Oxygen - chemistry
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Summary: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.
ISSN:1474-905X
1474-9092
DOI:10.1039/b9pp00091g