oxonol-v and 1-2-oleoylphosphatidylcholine

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.

Topics: Computer Simulation; Diffusion; Fluorescent Dyes; Isoxazoles; Kinetics; Liposomes; Mathematics; Membrane Potentials; Models, Biological; Phosphatidylcholines; Spectrometry, Fluorescence; Time Factors

The interaction of the dyes oxonol V and oxonol VI with unilamellar dioleoylphosphatidylcholine vesicles was investigated using a fluorescence stopped-flow technique. On mixing with the vesicles, both dyes exhibit an increase in their fluorescence, which occurs in two phases. According to the dependence of the reciprocal relaxation time on vesicle concentration, the rapid phase appears to be due to a second-order binding of the dye to the lipid membrane, which is very close to being diffusion-controlled. The slow phase is almost independent of vesicle concentration, and it is suggested that this may be due to a change in dye conformation or position within the membrane, possibly diffusion across the membrane to the internal monolayer. The response times of the dyes to a rapid jump in the membrane potential has also been investigated. Oxonol VI was found to respond to the potential change in less than 1 s, whereas oxonol required several minutes. This has been attributed to lower mobility of oxonol V within the lipid membrane.

Topics: Fluorescent Dyes; Isoxazoles; Kinetics; Liposomes; Mathematics; Membrane Potentials; Membranes; Models, Biological; Phosphatidylcholines; Spectrometry, Fluorescence