DETERMINATION OF DNA-PROTEIN AFFINITY CONSTANTS USING CHEMOMETRIC ANALYSIS OF TRYPTOPHAN FLUORESCENCE QUENCHING DATA

In this work we present a sensitive method to determine DNA protein af finity constants by analyzing tryptophan fluorescence quenching data. Chemometric analysis of the entire fluorescence emission spectra is us ed in combination with the equilibrium expression. This way all spectr al changes are taken into account, which considerably enhances the acc uracy compared to traditional methods where only the overall quenching is studied, In addition to the affinity constant, the concentrations in each sample and the spectral profiles of free and bound protein are determined.