| Abstrakt: |
Measurements of active encounters between molecules in native membranes containing ingredients, including proteins, are of prime importance. To estimate rare encounters in a high range of rate constants rate coefficients and distances between interacting molecules in membranes, a cascade of photochemical reactions for molecules diffusing in multilamellar liposomes was investigated. The sensitised cascade triplet cis–transphotoisomerisation of the excited stilbene involves the use of a triplet sensitiser Erythrosin B, a photochrome stilbene-derivative probe 4-dimethylamino-4′-aminostilbene exhibiting the phenomenon of trans–cisphotoisomerisation, and nitroxide radicals 5-doxyl stearic acid to quench the excited triplet state of the sensitiser. Measurement of the phosphorescence lifetime of Erythrosin B and the fluorescence enhancement of the stilbene-derivative photochrome probe, at various concentrations of the nitroxide probe, made it possible to calculate the quenching rate constant kq 1.1 × 1015cm2M−1s−1and the rate constant of the triplet–triplet energy transfer between the sensitiser and stilbene probe kT 1.0 × 1012cm2M−1s−1. These values, together with the data on diffusion rate constant, obtained by methods utilising various theoretical characteristic times of about seven orders of magnitude and the experimental rate constants of about five orders of magnitude, were found to be in good agreement with the advanced theory of diffusion-controlled reactions in two dimensions.1Because the characteristic time of the proposed cascade method is relatively large 0.1 s, it is possible to follow rare collisions between molecules and free radicals in model and biological membranes with a very sensitive fluorescence spectroscopy technique, using a relatively low concentration of probes. |
