Catalytic properties of recombinant dipeptidyl carboxypeptidase from Escherichia coli: a comparative study with angiotensin I-converting enzyme

Catalytic properties of recombinant dipeptidyl carboxypeptidase from Escherichia coli: a comparative study with angiotensin I-converting enzyme

Author Cunha, Carlos Eduardo L. Autor UNIFESP Google Scholar
Magliarelli, Helena de Fatima Autor UNIFESP Google Scholar
Paschoalin, Thaysa Autor UNIFESP Google Scholar
Nchinda, Aloysius T. Google Scholar
Lima, Jackson C. Autor UNIFESP Google Scholar
Juliano, Maria A. Autor UNIFESP Google Scholar
Paiva, Paulo B. Autor UNIFESP Google Scholar
Sturrock, Edward D. Google Scholar
Travassos, Luiz R. Autor UNIFESP Google Scholar
Carmona, Adriana K. Autor UNIFESP Google Scholar
Institution Universidade Federal de São Paulo (UNIFESP)
Univ Cape Town
Abstract Dipeptidyl carboxypeptidase from Escherichia coli (EcDcp) is a zinc metallopeptidase with catalytic properties closely resembling those of angiotensin I-converting enzyme (ACE). However, EcDcp and ACE are classified in different enzyme families (M3 and M2, respectively) due to differences in their primary sequences. We cloned and expressed EcDcp and studied in detail the enzyme's S(3) to S(1)' substrate specificity using positional-scanning synthetic combinatorial (PS-SC) libraries of fluorescence resonance energy transfer (FRET) peptides. These peptides contain ortho-aminobenzoic acid (Abz) and 2,4-dinitrophenyl (Dnp) as donor/acceptor pair. in addition, using FRET substrates developed for ACE [Abz-FRK(Dnp)P-OH, Abz-SDK(Dnp)P-OH and Abz-LFK(Dnp)-OH] as well as natural ACE substrates (angiotensin I, bradykinin, and Ac-SDKP-OH), we show that EcDcp has catalytic properties very similar to human testis ACE. EcDcp inhibition studies were performed with the ACE inhibitors captopril (K(i) = 3 nM) and lisinopril (K(i) = 4.4 mu M) and with two C-domain-selective ACE inhibitors, 5-S-5-benzamido-4-oxo-6-phenylhexanoyl-L-tryptophan (kAW; K(i) = 22.0 mu M) and lisinopril-Trp (K(i) = 0.8 nM). Molecular modeling was used to provide the basis for the differences found in the inhibitors potency. the phylogenetic relationship of EcDcp and related enzymes belonging to the M3 and M2 families was also investigated and the results corroborate the distinct origins of EcDcp and ACE.
Keywords angiotensin I-converting enzyme
dipeptidyl carboxypeptidase
molecular modeling
specificity studies
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Wellcome Trust, UK
National Research Foundation
Date 2009-09-01
Published in Biological Chemistry. Berlin: Walter de Gruyter & Co, v. 390, n. 9, p. 931-940, 2009.
ISSN 1431-6730 (Sherpa/Romeo, impact factor)
Publisher Walter de Gruyter & Co
Extent 931-940
Access rights Closed access
Type Article
Web of Science ID WOS:000269485600012

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