Light-enhanced inhibition of ouabain binding to digitalis receptor in rat brain and guinea pig heart by the food dye erythrosine.
Erythrosine (ERY) (FD & C red no. 3) inhibited specific binding of [3H]ouabain to rat brain homogenates with an IC50 of 23 microM in the dark and 1 microM in ordinary fluorescent light. Competition studies demonstrated the presence of two components, only one of which was affected by light. Lineweaver-Burk analysis indicated that ERY preferentially antagonizes [3H]ouabain binding at a high-affinity site in the light, whereas in the dark the dye inhibits binding in a manner qualitatively similar to inhibition by ouabain. Light enhancement of ERY potency occurred only when dye and tissue were present together in the incubation medium, pointing to participation of transient molecular species. However, neither superoxide dismutase nor catalase altered the effects of ERY in the light or dark, suggesting the absence of oxygen free radicals. When oxygen levels were raised, there was enhancement of inhibition by ERY at a high-affinity receptor accompanied by disappearance of [3H]ouabain binding at one of lower affinity. In contrast to brain, membranes from guinea pig heart showed only one binding site for [3H]ouabain, and antagonism by ERY at this site was markedly enhanced by light. Structural differences between classes of ouabain binding regions probably accounts for the discrimination exhibited by ERY in the presence of light and oxygen. Our findings also caution that metabolic transformation of this common food dye, light decomposition, or photoreaction with foodstuff may yield more toxic derivatives.
D000818:Animals; D001667:Binding, Competitive; D001921:Brain / Q000378:metabolism*; D002374:Catalase / Q000494:pharmacology; D004923:Erythrosine / Q000494:pharmacology*; D005452:Fluoresceins / Q000494:pharmacology*; D006168:Guinea Pigs; D066298:In Vitro Techniques; D007700:Kinetics; D008027:Light; D008297:Male; D009206:Myocardium / Q000378:metabolism*; D010042:Ouabain / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D011955:Receptors, Drug / Q000378:metabolism* / Q000528:radiation effects; D000254:Sodium-Potassium-Exchanging ATPase; D013482:Superoxide Dismutase / Q000494:pharmacology
D005452:Fluoresceins; D011955:Receptors, Drug; C027082:digitalis receptor; D010042:Ouabain; D002374:Catalase; D013482:Superoxide Dismutase; D000254:Sodium-Potassium-Exchanging ATPase; D004923:Erythrosine