Determination of carboxypeptidase A using N-acetyl-phenylalanyl-3-thiaphenylalanine as substrate: application to a direct serum assay

N-Acetyl-L-phenylalanyl-L-3-thiaphenylalanine has been shown to be a substrate for Carboxypeptidase A. Hydrolysis of the compound obeys Michaelis-Menten kinetics with a KM of 0.22 mM and a kcat of 6720 min-1 at 22 degrees C. A colorimetric assay, employing Ellman's reagent to detect the thiophenol released upon cleavage of the peptide, has been developed. The assay can be used for the direct determination of Carboxypeptidase A in serum.