AN IMPROVED ALGORITHM OF RESOLUTION OF FLUORESCENCE-SPECTRA INTO QUENCHER ACCESSIBILITY-ASSOCIATED COMPONENTS

A novel algorithm (QUENCH) is described that resolves fluorescence spe ctra into components differing in the accessibility (the Stern-Volmer constants) for small quencher molecules. In contrast to the known algo rithms, QUENCH first evaluates Stern-Volmer constants of the component s using the total set of intensities in spectra measured at several qu encher concentrations and then calculates contributions of each compon ent in the emission intensities at every wavelength value, The compone nt spectra, revealed with QUENCH in the tryptophan fluorescence of rib osomal S7 protein, are very similar to the log-normal components calcu lated using the SIMS program (Abornev, S. M. and E. A. Burstein Mol. B iol. [Engl. transl.] 26, 890-897 1992). The QUENCH method is applicabl e for the component resolution of fluorescence spectra of any liquid s ystem containing fluorophores of different accessibility to small quen chers.