title: Photophysics and photochemistry of dyes bound to human serum albumin are determined by the dye localization.
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.
authors.affiliation: Facultad de Química, Departamento de Química Física, Pontificia Universidad Católica de Chile, Santiago, Chile.
authors.forename: Emilio
authors.initials: E
authors.lastname: Alarcón
authors.forename: Ana Maria
authors.initials: AM
authors.lastname: Edwards
authors.forename: Alexis
authors.initials: A
authors.lastname: Aspee
authors.forename: Faustino E
authors.initials: FE
authors.lastname: Moran
authors.forename: Claudio D
authors.initials: CD
authors.lastname: Borsarelli
authors.forename: Eduardo A
authors.initials: EA
authors.lastname: Lissi
authors.forename: Danilo
authors.initials: D
authors.lastname: Gonzalez-Nilo
authors.forename: Horacio
authors.initials: H
authors.lastname: Poblete
authors.forename: J C
authors.initials: JC
authors.lastname: Scaiano
mesh_terms: D001665:Binding Sites; D004396:Coloring Agents / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008751:Methylene Blue / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D055668:Photochemical Processes; D011487:Protein Conformation; D012395:Rose Bengal / Q000737:chemistry / Q000378:metabolism; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism; D026082:Singlet Oxygen / Q000737:chemistry
chemical_list: D004396:Coloring Agents; D012709:Serum Albumin; D026082:Singlet Oxygen; D012395:Rose Bengal; D008751:Methylene Blue