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

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

Relationships between angiotensin I converting enzyme gene polymorphism, plasma levels, and diabetic retinal and renal complications

Diabetes ◽  
1994 ◽  
Vol 43 (3) ◽  
pp. 384-388 ◽  
Author(s):  
M. Marre ◽  
P. Bernadet ◽  
Y. Gallois ◽  
F. Savagner ◽  
T. T. Guyene ◽  
...  
Keyword(s):  
Gene Polymorphism ◽  
Plasma Levels ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Enzyme Gene ◽  
Angiotensin I Converting Enzyme ◽  
Renal Complications

Angiotensin I-Converting Enzyme (ACE) Inhibitory Activity of Elk (Cervus elaphus) Velvet Antler

2005 ◽  
Vol 10 (3) ◽  
pp. 239-243 ◽  
Author(s):  
Rohan Karawita ◽  
Pyo-Jam park ◽  
Nalin Siriwardhana ◽  
Byong-Tae Jeon ◽  
Sang-Ho Moon ◽  
...  
Keyword(s):  
Cervus Elaphus ◽  
Inhibitory Activity ◽  
Converting Enzyme ◽  
Angiotensin I ◽  
Ace Inhibitory Activity ◽  
Angiotensin I Converting Enzyme ◽  
Velvet Antler ◽  
Ace Inhibitory
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