The two homologous Domains of human angiotensin I-converting enzyme are both catalytically active

Lei Wei, François Alhenc-Gelas, Pierre Corvol, Eric Clauser

Research output: Contribution to journalArticle

320 Citations (Scopus)

Abstract

Molecular cloning of human endothelial angiotensin I-converting enzyme (kininase II; EC 3. 4. 15. 1) (ACE) has recently shown that the enzyme contains two large homologous domains (called here the N and C domains), each bearing a putative active site, identified by sequence comparisons with the active sites of other zinc metallopeptidases. However, the previous experiments with zinc or competitive ACE inhibitors suggested a single active site in ACE. To establish whether both domains of ACE are enzymatically active, a series of ACE mutants, each containing only one intact domain, were constructed by deletion or point mutations of putative critical residues of the other domain, and expressed in heterologous Chinese hamster ovary cells. Both domains are enzymatically active and cleave the C-terminal dipeptide of hippuryl-His-Leu or angiotensin I. Moreover, both domains have an absolute zinc requirement for activity, are activated by chloride and are sensitive to competitive ACE inhibitors, and appear to function independently. However, the two domains display different catalytic constants and different patterns of chloride activation. At high chloride concentrations, the C domain hydrolyzes the two substrates tested faster than does the N domain. His-361,365 and His-959,963 are established as essential residues in the N and C domains, respectively, most likely involved in zinc binding, and Glu-362 in the N domain and Glu-960 in the C domain are essential catalytic residues. These observations provide strong evidence that ACE possesses two independent catalytic domains and suggest that they may have different functions.

Original languageEnglish (US)
Pages (from-to)9002-9008
Number of pages7
JournalJournal of Biological Chemistry
Volume266
Issue number14
StatePublished - 1991
Externally publishedYes

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Peptidyl-Dipeptidase A
Zinc
Catalytic Domain
Chlorides
Angiotensin-Converting Enzyme Inhibitors
Bearings (structural)
Angiotensin I
Dipeptides
Cloning
Metalloproteases
Sequence Deletion
Molecular Cloning
Cricetulus
Point Mutation
Chemical activation
Cells
Ovary
Substrates
Enzymes
Experiments

ASJC Scopus subject areas

  • Biochemistry

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The two homologous Domains of human angiotensin I-converting enzyme are both catalytically active. / Wei, Lei; Alhenc-Gelas, François; Corvol, Pierre; Clauser, Eric.

In: Journal of Biological Chemistry, Vol. 266, No. 14, 1991, p. 9002-9008.

Research output: Contribution to journalArticle

Wei, Lei ; Alhenc-Gelas, François ; Corvol, Pierre ; Clauser, Eric. / The two homologous Domains of human angiotensin I-converting enzyme are both catalytically active. In: Journal of Biological Chemistry. 1991 ; Vol. 266, No. 14. pp. 9002-9008.
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