Catalytic Properties of the Two Active Sites of Angiotensin I-Converting Enzyme on the Cell Surface

1995 ◽  
Vol 211 (2) ◽  
pp. 528-534 ◽  
Author(s):  
E. Jaspard ◽  
F. Alhencgelas
Keyword(s):  
Cell Surface ◽  
Active Sites ◽  
Catalytic Properties ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Angiotensin I Converting Enzyme

scholarly journals Cell Surface Localization of Proteolysis of Human Endothelial Angiotensin I-converting Enzyme

1995 ◽  
Vol 270 (48) ◽  
pp. 28962-28969 ◽  
Author(s):  
Véronique Beldent ◽  
Annie Michaud ◽  
Christophe Bonnefoy ◽  
Marie-Thérèse Chauvet ◽  
Pierre Corvol
Keyword(s):  
Cell Surface ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Angiotensin I Converting Enzyme ◽  
Surface Localization ◽  
Cell Surface Localization

scholarly journals Angiotensin I-Converting Enzyme Gln1069Arg Mutation Impairs Trafficking to the Cell Surface Resulting in Selective Denaturation of the C-Domain

PLoS ONE ◽  
2010 ◽  
Vol 5 (5) ◽  
pp. e10438 ◽  
Author(s):  
Sergei M. Danilov ◽  
Sergey Kalinin ◽  
Zhenlong Chen ◽  
Elena I. Vinokour ◽  
Andrew B. Nesterovitch ◽  
...  
Keyword(s):  
Cell Surface ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Angiotensin I Converting Enzyme

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

2009 ◽  
Vol 390 (9) ◽  
Author(s):  
Carlos Eduardo L. Cunha ◽  
Helena de Fátima Magliarelli ◽  
Thaysa Paschoalin ◽  
Aloysius T. Nchinda ◽  
Jackson C. Lima ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Ace Inhibitors ◽  
Catalytic Properties ◽  
Resonance Energy ◽  
Human Testis ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Acceptor Pair ◽  
Angiotensin I Converting Enzyme ◽  
Dipeptidyl Carboxypeptidase

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 S3 to S1′ 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 μm) and with two C-domain-selective ACE inhibitors, 5-S-5-benzamido-4-oxo-6-phenylhexanoyl-L-tryptophan (kAW; K i=22.0 μm) 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.

scholarly journals Drosophila melanogaster angiotensin I-converting enzyme expressed in Pichia pastoris resembles the C domain of the mammalian homologue and does not require glycosylation for secretion and enzymic activity

1996 ◽  
Vol 318 (1) ◽  
pp. 125-131 ◽  
Author(s):  
Tracy A. WILLIAMS ◽  
Annie MICHAUD ◽  
Xavier HOUARD ◽  
Marie-Thérèse CHAUVET ◽  
Florent SOUBRIER ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
Pichia Pastoris ◽  
Ace Inhibitors ◽  
Active Sites ◽  
Chloride Concentration ◽  
Enzymic Activity ◽  
Expression Level ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Angiotensin I Converting Enzyme

Drosophila melanogaster angiotensin I-converting enzyme (AnCE) is a secreted single-domain homologue of mammalian angiotensin I-converting enzyme (ACE) which comprises two domains (N and C domains). In order to characterize in detail the enzymic properties of AnCE and to study the influence of glycosylation on the secretion and enzymic activity of this enzyme, we overexpressed AnCE (expression level, 160 mg/l) and an unglycosylated mutant (expression level, 43 mg/l) in the yeast Pichia pastoris. The recombinant enzyme was apparently homogeneous on SDS/PAGE without purification and partial deglycosylation demonstrated that all three potential sites for N-linked glycosylation were occupied by oligosaccharide chains. Each N-glycosylation sequence (Asn-Xaa-Ser/Thr) was disrupted by substituting a glutamine for the asparagine residue at amino acid positions 53, 196 and 311 by site-directed mutagenesis to produce a single mutant. Expression of the unglycosylated mutant in Pichia produced a secreted catalytically active enzyme (AnCEΔCHO). This mutant displayed unaltered kinetics for the hydrolyses of hippuryl-His-Leu, angiotensin I and N-acetyl-Ser-Asp-Lys-Pro (AcSDKP) and was equally sensitive to ACE inhibitors compared with wild-type AnCE. However, AnCEΔCHO was less stable, displaying a half-life of 4.94 h at 37 °C, compared with AnCE which retained full activity under the same conditions. Two catalytic criteria demonstrate the functional resemblance of AnCE with the human ACE C domain: first, the kcat/Km of AcSDKP hydrolysis and secondly, the kcat/Km and optimal chloride concentration for hippuryl-His-Leu hydrolysis. A range of ACE inhibitors were far less potent towards AnCE compared with the human ACE domains, except for captopril which suggests an alternative structure in AnCE corresponding to the region of the S1 subsite in the human ACE active sites.

SPECIFICITY EVALUATION OF THE TWO ACTIVE SITES OF ANGIOTENSIN I CONVERTING ENZYME USING COMBINATORIAL FLUORESCENT-QUENCHED PEPTIDE LIBRARIES

2004 ◽  
Vol 22 (Suppl. 1) ◽  
pp. S176
Author(s):  
Patricia Alessandra Bersanetti ◽  
M. C.C. Andrade ◽  
M. A. Juliano ◽  
D. E. Casarini ◽  
E. D. Sturrock ◽  
...  
Keyword(s):  
Active Sites ◽  
Peptide Libraries ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Angiotensin I Converting Enzyme

Zinc binding in peptide models of angiotensin-I converting enzyme active sites studied through1H-NMR and chemical shift perturbation mapping

Biopolymers ◽  
2003 ◽  
Vol 69 (2) ◽  
pp. 244-252 ◽  
Author(s):  
Athanassios S. Galanis ◽  
Georgios A. Spyroulias ◽  
Roberta Pierattelli ◽  
Andreas Tzakos ◽  
Anastassios Troganis ◽  
...  
Keyword(s):  
Chemical Shift ◽  
Active Sites ◽  
Zinc Binding ◽  
Chemical Shift Perturbation ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Angiotensin I Converting Enzyme ◽  
Enzyme Active Sites ◽  
Peptide Models
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