pH-dependent interaction of rhodopsin with cyanidin-3-glucoside. 2. Functional aspects.
Anthocyanins are a class of phytochemicals that confer color to flowers, fruits, vegetables and leaves. They are part of our regular diet and serve as dietary supplements because of numerous health benefits, including improved vision. Recent studies have shown that the anthocyanin cyanidin-3-O-glucoside (C3G) increased regeneration of the dim-light photoreceptor rhodopsin (Matsumoto et al. [2003] J. Agric. Food Chem., 51, 3560-3563). In an accompanying study (Yanamala et al. [2009] Photochem. Photobiol.), we show that C3G directly binds to rhodopsin in a pH-dependent manner. In this study, we investigated the functional consequences of C3G binding to rhodopsin. As observed previously in rod outer segments, regeneration of purified rhodopsin in detergent micelles is also accelerated in the presence of C3G. Thermal denaturation and stability studies using circular dichroism, fluorescence and UV/visible absorbance spectroscopy show that C3G exerts a destabilizing effect on rhodopsin structure while it only modestly alters G-protein activation and the rates at which the light-activated Metarhodopsin II state decays to opsin and free retinal. These results indicate that the mechanism of C3G-enhanced regeneration may be based on changes in opsin structure promoting access to the retinal binding pocket.
Tirupula
Kalyan C
KC
Department of Structural Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
Balem
Fernanda
F
Yanamala
Naveena
N
Klein-Seetharaman
Judith
J
D000818:Animals; D000872:Anthocyanins / Q000737:chemistry* / Q000378:metabolism*; D002417:Cattle; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism; D005960:Glucosides / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D011485:Protein Binding; D017433:Protein Structure, Secondary; D012243:Rhodopsin / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*
D000872:Anthocyanins; D005960:Glucosides; C462279:cyanidin-3-O-beta-glucopyranoside; D012243:Rhodopsin; D019204:GTP-Binding Proteins