Interaction of second and third domains of Japanese quail ovomucoid with ten mammalian trypsins.

Second and third domains were prepared from Japanese quail ovomucoid and association equilibrium constants, Kas, were measured at 25 degreesC and pH 8 for these domains with trypsins from ten mammalian species: cat, cow, dog, guinea pig, hog, horse, man, rabbit, rat, and sheep. The values ranged from 108 M-1 to 1010 M-1 for the second domain-trypsin associations and from 106 M-1 to 108 M-1 for the third domain-trypsin associations. Changes in Ka values for the interactions between the trypsins and each domain are attributed to slight changes in surface conformation caused by the residue changes in the inhibitor-binding region other than the S1 pocket of the trypsin species. The representative of such residue changes is assumed to be the one observed at residue 217 of trypsin molecule. Concerning each trypsin, the Ka value with the second domain was always higher than that with the third domain. However, the ratios between the two equilibrium constants varied from 3 to 60 depending upon trypsin species. This means that amino acid changes in enzyme-contact residues other than the P1 site of the Kazal-type inhibitor can make it possible to recognize even a slight difference in inhibitor-binding surface among the enzymes with the same S1 pocket and highly similar overall three-dimensional structure.