A fluorometric assay for angiotensin-converting enzyme activity |
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| Authors: | M S Kapiloff S M Strittmatter L D Fricker S H Snyder |
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| Affiliation: | 1. Department of Neuroscience, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, Maryland 21205 USA;2. Department of Pharmacology and Experimental Therapeutics, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, Maryland 21205 USA;3. Department of Psychiatry and Behavioral Sciences, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Baltimore, Maryland 21205 USA |
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| Abstract: | 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. |
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| Keywords: | fluorometric enzyme assay dansyl-tripeptides sarcoidosis hippuryl-histidylleucine |
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