[ANSA analysis. V. Secondary specificity of peptidases to the structure of detected groups of aminonaphthalenesulfamide chromogenic substrates].

Mixtures of competing aminonaphthalenesulfonamide (ANSA) substrates can be used for identifying secondary substrate specificity of peptidases and their mixtures. All substrates in the mixture had identical amino acid residues; however, cleaved ANSA differed in chromatographic mobilities due to differences in the substituent in the sulfonamide residue. ANSA generated by limited proteolysis of the substrate mixture by the enzyme preparation were detected chromatographically. The chromatogram obtained (the ANSA spectrum) was characteristic of each enzyme or enzyme-containing preparation. Using arginyl-ANSA mixtures, informative ANSA spectra suitable for enzyme identification were obtained. The correlations between the structure of the substituents in the SONR1R2 group of ANSA and the efficiency of substrate hydrolysis were studied. The method was used to identify snake venoms.