A fluorometric assay for angiotensin-converting enzyme activity

Michael S. Kapiloff; Stephen M. Strittmatter; Lloyd D. Fricker; Solomon H. Snyder

doi:10.1016/0003-2697(84)90167-2

A fluorometric assay for angiotensin-converting enzyme activity

Michael S. Kapiloff, Stephen M. Strittmatter, Lloyd D. Fricker, Solomon H. Snyder

Research output: Contribution to journal › Article › peer-review

17 Scopus citations

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 dansyl-phenylalanyl-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.

Original language	English (US)
Pages (from-to)	293-302
Number of pages	10
Journal	Analytical Biochemistry
Volume	140
Issue number	1
DOIs	https://doi.org/10.1016/0003-2697(84)90167-2
State	Published - Jul 1984
Externally published	Yes

Keywords

dansyl-tripeptides
fluorometric enzyme assay
hippuryl-histidylleucine
sarcoidosis

ASJC Scopus subject areas

Biophysics
Biochemistry
Molecular Biology
Cell Biology

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10.1016/0003-2697(84)90167-2

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title = "A fluorometric assay for angiotensin-converting enzyme activity",

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 dansyl-phenylalanyl-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.",

keywords = "dansyl-tripeptides, fluorometric enzyme assay, hippuryl-histidylleucine, sarcoidosis",

author = "Kapiloff, {Michael S.} and Strittmatter, {Stephen M.} and Fricker, {Lloyd D.} and Snyder, {Solomon H.}",

note = "Funding Information: We thank Dawn C. Dodson for manuscript preparation and Patricia W. Salevan of the American Red Cross Blood Services, Chesapeake region, for assistance in obtaining human serum samples. This work was supported by USPHS Grants MH-18501, DA-00266, NS16375; RSA Award DA-00074 to S.H.S.; MSTP Grant GM-07309 to S.M.S.; and a grant of the M&night Foundation.",

year = "1984",

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doi = "10.1016/0003-2697(84)90167-2",

language = "English (US)",

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pages = "293--302",

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AU - Kapiloff, Michael S.

AU - Strittmatter, Stephen M.

AU - Fricker, Lloyd D.

AU - Snyder, Solomon H.

N1 - Funding Information: We thank Dawn C. Dodson for manuscript preparation and Patricia W. Salevan of the American Red Cross Blood Services, Chesapeake region, for assistance in obtaining human serum samples. This work was supported by USPHS Grants MH-18501, DA-00266, NS16375; RSA Award DA-00074 to S.H.S.; MSTP Grant GM-07309 to S.M.S.; and a grant of the M&night Foundation.

PY - 1984/7

Y1 - 1984/7

N2 - 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 dansyl-phenylalanyl-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.

AB - 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 dansyl-phenylalanyl-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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KW - fluorometric enzyme assay

KW - hippuryl-histidylleucine

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A fluorometric assay for angiotensin-converting enzyme activity

Abstract

Keywords

ASJC Scopus subject areas

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