A fluorometric assay for angiotensin-converting enzyme activity.
A simple and sensitive assay for angiotensin-converting enzyme (ACE; EC 3.4.15.1) activity has been developed which employs fluorescently labeled tripeptides. ACE hydrolyzes dansylphenylalanyl-arginyl-tryptophan or dansyl-phenylalanyl-arginyl-phenylalanine, liberating dansyl-phenylalanine and a dipeptide. Dansyl-phenylalanine partitions quantitatively into chloroform, whereas the substrates are virtually insoluble in chloroform. This allows rapid measurement of ACE activity with high signal-to-noise ratios even when microliter aliquots of human serum are assayed. Inhibition studies of the dansyl-tripeptide cleaving activity of human serum and rat lung, the identity of the products of enzyme action, and the regional distribution of enzyme activity among rat tissues demonstrate that only ACE cleaves these substrates under the conditions employed here. This assay may be useful for the clinical measurement of human serum ACE activity and for research investigations of ACE from a variety of tissues.
D000818:Animals; D002855:Chromatography, Thin Layer; D003619:Dansyl Compounds / Q000138:chemical synthesis / Q000378:metabolism; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D006801:Humans; D007700:Kinetics; D008168:Lung / Q000201:enzymology; D008297:Male; D009842:Oligopeptides / Q000138:chemical synthesis / Q000378:metabolism; D007703:Peptidyl-Dipeptidase A / Q000097:blood*; D051381:Rats; D011919:Rats, Inbred Strains; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D003619:Dansyl Compounds; D005456:Fluorescent Dyes; D009842:Oligopeptides; D007703:Peptidyl-Dipeptidase A