CARBOXYPEPTIDASE-A HYDROLYZES BENZOYLGLYCYL-HISTIDYL-LEUCINE BUT NOT FURYLACRYLOYL-PHENYLALANYL-GLYCYL-GLYCINE, 2 USUAL SUBSTRATES FOR ANGIOTENSIN-I-CONVERTING ENZYME

We compared angiotensin I-converting enzyme (ACE) and carboxypeptidase A (CPA), two zinc metallopeptidases, for the hydrolysis of the usual ACE synthetic substrate benzoylglycyl-histidyl-leucine (HHL) investiga ting the possible interference by CPA in the determination of ACE acti vity in biological fluids. Both purified enzymes hydrolyse HHL in a ra diochemical assay with the same optimal pH, a characteristic divalent metal requirement, a close similar behavior against inhibitors of othe r metallopeptidases, such as enkephalinase and kininase I, and the inv olvement of arginine and lysine residues in their active site. Convers ely, CPA does not show the other catalytic properties of ACE, i.e. chl oride dependence, low K-m for HHL, inhibition by specific synthetic AC E inhibitors and antibody, also hydrolysis of the other ACE substrate furyl-acryloylphenylalanyl-glycyl-glycine (FAPGG). We advise the use o f ACE inhibitors to validate ACE measurement with HHL or, alternativel y, FAPGG, which is a more specific substrate for ACE, must be preferre d, although the poor sensitivity of the spectro-photometric assay with this substrate limits its use to blood samples.