Binding and diffusion kinetics of the interaction of a hydrophobic potential-sensitive dye with lipid vesicles

The interaction of the dye oxonol V with unilamellar dioleoylphosphatidylcholine vesicles has previously been investigated using a fluorescence stopped-flow technique. It has been found that the most suitable mathematical description of the equilibrium and kinetic data is obtained by assuming the presence of saturable dye binding sites in both monolayers of the vesicle membrane and a potential-dependent diffusion across the membrane interior between these two classes of sites. A kinetic model is presented which takes into account the degree of saturation of the binding sites, the degree of fluorescence quenching within the membrane, and the production of an electrical potential gradient across the membrane interior by the binding of the negatively charged dye. The model successfully predicts the time course of the fluorescence change due to binding and diffusion over the complete range of dye and vesicle concentrations as well as the fluorescence response of the dye to changing membrane potential.