A Fluorometric Method of Measuring Carboxypeptidase Activities for Angiotensin II and Apelin-13

Pan Liu, Jan Wysocki, Peter Serfozo, Minghao Ye, Tomokazu Souma, Daniel Batlle, Jing Jin*

*Corresponding author for this work

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Degradation of the biologically potent octapeptide angiotensin Ang II-(1-8) is mediated by the activities of several peptidases. The conversion of Ang II to the septapeptide Ang-(1-7) is of particular interest as the latter also confers organ protection. The conversion is catalyzed by angiotensin-converting enzyme 2 and other enzymes that selectively cleave the peptide bond between the proline and the phenylalanine at the carboxyl terminus of Ang II. The contribution of various enzyme activities that collectively lead to the formation of Ang-(1-7) from Ang II, in both normal conditions and in disease states, remains only partially understood. This is largely due to the lack of a reliable and sensitive method to detect these converting activities in complex samples, such as blood and tissues. Here, we report a fluorometric method to measure carboxypeptidase activities that cleave the proline-phenylalanine dipeptide bond in Ang II. This method is also suitable for measuring the conversion of apelin-13. The assay detects the release of phenylalanine amino acid in a reaction with the yeast enzyme of phenylalanine ammonia lyase (PAL). When used in cell and mouse organs, the assay can robustly measure endogenous Ang II and apelin-13-converting activities involved in the renin-angiotensin and the apelinergic systems, respectively.

Original languageEnglish (US)
Article number45473
JournalScientific reports
Volume7
DOIs
StatePublished - Apr 5 2017

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Carboxypeptidases
Phenylalanine
Angiotensin II
Proline
Enzymes
Phenylalanine Ammonia-Lyase
Dipeptides
Renin-Angiotensin System
Peptide Hydrolases
Yeasts
Amino Acids
Peptides
apelin-13 peptide

ASJC Scopus subject areas

  • General

Cite this

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title = "A Fluorometric Method of Measuring Carboxypeptidase Activities for Angiotensin II and Apelin-13",
abstract = "Degradation of the biologically potent octapeptide angiotensin Ang II-(1-8) is mediated by the activities of several peptidases. The conversion of Ang II to the septapeptide Ang-(1-7) is of particular interest as the latter also confers organ protection. The conversion is catalyzed by angiotensin-converting enzyme 2 and other enzymes that selectively cleave the peptide bond between the proline and the phenylalanine at the carboxyl terminus of Ang II. The contribution of various enzyme activities that collectively lead to the formation of Ang-(1-7) from Ang II, in both normal conditions and in disease states, remains only partially understood. This is largely due to the lack of a reliable and sensitive method to detect these converting activities in complex samples, such as blood and tissues. Here, we report a fluorometric method to measure carboxypeptidase activities that cleave the proline-phenylalanine dipeptide bond in Ang II. This method is also suitable for measuring the conversion of apelin-13. The assay detects the release of phenylalanine amino acid in a reaction with the yeast enzyme of phenylalanine ammonia lyase (PAL). When used in cell and mouse organs, the assay can robustly measure endogenous Ang II and apelin-13-converting activities involved in the renin-angiotensin and the apelinergic systems, respectively.",
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A Fluorometric Method of Measuring Carboxypeptidase Activities for Angiotensin II and Apelin-13. / Liu, Pan; Wysocki, Jan; Serfozo, Peter; Ye, Minghao; Souma, Tomokazu; Batlle, Daniel; Jin, Jing.

In: Scientific reports, Vol. 7, 45473, 05.04.2017.

Research output: Contribution to journalArticle

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AU - Liu, Pan

AU - Wysocki, Jan

AU - Serfozo, Peter

AU - Ye, Minghao

AU - Souma, Tomokazu

AU - Batlle, Daniel

AU - Jin, Jing

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