Novel fluorescence-photochrome labeling method in the study of biomembrane dynamics

A novel photochrome-fluorescence method (PFLM) based on monitoring fluorescence parameters and kinetics of photochrome photoisomerization of para-substituted stilbenes (PSS) has been proposed. It was shown that PSS exhibits fluorescence characteristics which are similar to ones of typical membrane fluorescence probes such as diphenylhexatriene (DPH). A study of kinetics of PSS trans-cis and cis-trans photoisomerization makes it possible to estimate, under certain conditions, the rotational correlation time of the stilbene fragments in the excited state of PSS for the fixed angle 180 degrees. In viscous media this process is a rate-determining stage. Taken together, the both techniques, fluorescence and photochrome, make it possible to establish a detailed mechanism and measure quantitative parameters of stilbene probe (PSS) mobility in a membrane. The PFLM was applied to the study of E. coli membrane dynamics.