Phosphonodipeptides containing (2-aminoethyl)phosphonic acid (ciliatine): transition state analogue inhibitors of carboxypeptidase A

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Abstract The phosphonodipeptides having a structure of Cbz-NH(CH2)2PO(O−Li+)-NHCHRCO2−Li+ (Cbz = carbobenzyloxyl; amino acid residues Gly, l -Ala and l -Phe) were found to inhibit competitively the hydrolysis of Cbz-Gly-Gly- l Phe-OH by carboxypeptidase A. The compound having an l -phenylalanine residue was especially effective, giving Ki values of about 10−8 and 10−9 M at pH 7.5 and 6.0 (25°C), respectively. The kinetics suggested that (i) an ionized P-O− group of the inhibitors with a tetrahedral phosphorus atom forms a coordinating bond with a zinc ion of the enzyme and (ii) the amino acid side-chains of the inhibitors are accommodated in the hydrophoic pocket of the enzyme. The inhibitory activity of the phosphonodipeptides was explained by a transition state analog mechanism.

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