An improved algorithm of resolution of fluorescence spectra into quencher accessibility-associated components

A novel algorithm (QUENCH) is described that resolves fluorescence spectra into components differing in the accessibility (the Stern-Volmer constants) for small quencher molecules. In contrast to the known algorithms, QUENCH first evaluates Stern-Volmer constants of the components using the total set of intensities in spectra measured at several quencher concentrations and then calculates contributions of each component in the emission intensities at every wavelength value, The component spectra, revealed with QUENCH in the tryptophan fluorescence of ribosomal S7 protein, are very similar to the log-normal components calculated using the SIMS program (Abornev, S. M. and E. A. Burstein Mol. Biol. [Engl. transl.] 26, 890-897 1992). The QUENCH method is applicable for the component resolution of fluorescence spectra of any liquid system containing fluorophores of different accessibility to small quenchers.