Interaction between trypsin-like enzyme from Streptomyces erythraeus and Japanese quail ovomucoid.

We examined the interaction between a trypsin-like enzyme from Streptomyces erythraeus (TLE-Se) and Japanese quail ovomucoid (QO) as a model of the interaction between a serine protease and a Kazal-type inhibitor. Both the second domain (Domain II) and the third domain (Domain III) of QO inhibited TLE-Se. But there was a great difference in the apparent association rate constant (kappa a) between Domain II and Domain III at pH 8.0 (the optimum pH of the enzymatic activity), and Domain III associated with TLE-Se about 10(5) times faster than Domain II. The pH dependency of kappa a for Domain III and TLE-Se was maximum around pH 8, but in the case of Domain II and TLE-Se the maximum was around pH 4. This suggested that some acidic amino acid residues had some influence upon the association of Domain II with TLE-Se. When we examined the interaction between TLE-Se and amidated Domain II in which most of the aspartic and glutamic acids were amidated for conversion into asparagine and glutamine with NH4Cl and carbodiimide, the pH dependence of kappa a greatly differed from that obtained for TLE-Se and native Domain II, and an almost constant kappa a value (10(3)-10(5) times higher than that of native Domain II) was exhibited between pH 5 and 8. At the same time, the dissociation constant at pH 8.0 became about 10(-2) times smaller, and thus the affinity between TLE-Se and amidated Domain II became stronger. These observations suggest that the electrostatic effect derived from the repulsion between the minus charges which are located at contact area sites of both molecules control the reactivity in the interaction between protease and a protein protease inhibitor.