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 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.
Alarcón
Emilio
E
Facultad de Química, Departamento de Química Física, Pontificia Universidad Católica de Chile, Santiago, Chile.
Edwards
Ana Maria
AM
Aspee
Alexis
A
Moran
Faustino E
FE
Borsarelli
Claudio D
CD
Lissi
Eduardo A
EA
Gonzalez-Nilo
Danilo
D
Poblete
Horacio
H
Scaiano
J C
JC
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
D004396:Coloring Agents; D012709:Serum Albumin; D026082:Singlet Oxygen; D012395:Rose Bengal; D008751:Methylene Blue