Reaction of azapeptides with chymotrypsin-like enzymes. New inhibitors and active site titrants for chymotrypsin A alpha, subtilisin BPN', subtilisin Carlsberg, and human leukocyte cathepsin G.

Abstract

A series of new azapeptide p-nitrophenyl esters containing a variety of P1 aza-amino acid residues have been synthesized, and the reaction of these azapeptides with chymotrypsin A alpha, subtilisin BPN', subtilisin Carlsberg, and human leukocyte cathepsin G at pH 4-7 has been studied. These azapeptides were found to be very useful as active site titrants and inhibitors of serine proteases with chymotrypsin-like specificity. Stable acyl derivatives of serine proteases are formed in the reaction with azapeptides and can be used for future crystallographic investigations. The effects of changing the nature of the P1' leaving group (-ONp, -OPh, -OCH2CF3, -OEt) for these azapeptides was also investigated. N-Acetyl-L-alanyl-L-alanyl-alpha-azanorleucine p-nitrophenyl ester can be used as an active site titrant for human leukocyte cathepsin G and N-acetyl-L-alanyl-alpha-azaphenylalanine p-nitrophenyl ester is a suitable titrant for chymotrypsin A alpha, subtilisins, or cathepsin G.