Identification of two active site residues in human angiotensin I-converting enzyme.

Angiotensin I-converting enzyme (ACE) contains two zinc-dependent catalytic domains (N and C domains) each bearing the motif HEXXH where the two histidines form two of the three amino acid zinc ligands. Sequence alignment of each ACE domain with other zinc metalloproteases, indicates a glutamate residues which putatively constitutes the third zinc ligand and an aspartate residue which may form an indirect zinc interaction. We investigated the functional roles of the glutamate and aspartate residues in the ACE C domain (Glu987 and Asp991) using a cDNA encoding an inactive N domain. We mutated Glu987 to aspartate (E987D) or valine (E987V) and Asp991 to glutamate (D991E) or alanine (D991A). Catalytically active mutants (E987D, D991E and D991A) exhibited similar Km values for hippuryl-His-Leu compared to non-mutated C domain. E987D displayed a 300-fold decrease in kcat and a 25-fold reduction in sensitivity to the ACE inhibitor trandolaprilat, whose binding is zinc-dependent. E987V was catalytically inactive and did not bind [3H]trandolaprilat. D991E and D991A exhibited a 3.8- and 22-fold decrease in kcat, respectively, and the Ki' values for trandolaprilat were increased 8- and 29-fold. These results provide strong evidence that Glu987 constitutes the third zinc ligand in the ACE C domain and suggest a role for Asp991 in positioning the C domain active site zinc ion.

• Publication year

  1994

• Venue

  Journal of Biological Chemistry

• Publication date

  1994-11-25

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(18)43897-5PMID 7961923
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• The metallobiochemistry of zinc enzymes.

  2006cited by this paper
• Mutations in two specific residues of testicular angiotensin-converting enzyme change its catalytic properties.

  1993cited by this paper
• Families of metalloendopeptidases and their relationships

  1992cited by this paper
• A comparison of the zinc contents and substrate specificities of the endothelial and testicular forms of porcine angiotensin converting enzyme and the preparation of isoenzyme-specific antisera.

  1992cited by this paper
• The two homologous domains of human angiotensin I-converting enzyme interact differently with competitive inhibitors.

  1992cited by this paper
• The two homologous domains of human angiotensin I-converting enzyme are both catalytically active.

  1991cited by this paper
• Angiotensin-converting enzyme: zinc- and inhibitor-binding stoichiometries of the somatic and testis isozymes.

  1991cited by this paper
• Identification of glutamic acid 646 as a zinc-coordinating residue in endopeptidase-24.11.

  1991cited by this paper
• Expression and characterization of recombinant human angiotensin I-converting enzyme. Evidence for a C-terminal transmembrane anchor and for a proteolytic processing of the secreted recombinant and plasma enzymes.

  1991cited by this paper
• Active-site zinc ligands and activated H2O of zinc enzymes.

  1990cited by this paper
• Molecular cloning of human testicular angiotensin-converting enzyme: the testis isozyme is identical to the C-terminal half of endothelial angiotensin-converting enzyme.

  1989cited by this paper
• The testicular transcript of the angiotensin I‐converting enzyme encodes for the ancestral, non‐duplicated form of the enzyme

  1989cited by this paper
• Angiotensin-converting enzyme inhibition, anti-hypertensive activity and hemodynamic profile of trandolapril (RU 44570).

  1988cited by this paper
• Two putative active centers in human angiotensin I-converting enzyme revealed by molecular cloning.

  1988cited by this paper
• Amino acid sequence of rabbit kidney neutral endopeptidase 24.11 (enkephalinase) deduced from a complementary DNA.

  1987cited by this paper
• Replacement of insulin receptor tyrosine residues 1162 and 1163 compromises insulin-stimulated kinase activity and uptake of 2-deoxyglucose.

  1986cited by this paper
• Rapid and efficient site-specific mutagenesis without phenotypic selection.

  1985cited by this paper
• Inhibition of rabbit lung angiotensin-converting enzyme by N alpha-[(S)-1-carboxy-3-phenylpropyl]L-alanyl-L-proline and N alpha-[(S)-1-carboxy-3-phenylpropyl]L-lysyl-L-proline.

  1985cited by this paper
• Measurement of human converting enzyme level by direct radioimmunoassay.

  1983cited by this paper
• Molecular and catalytic properties of rabbit testicular dipeptidyl carboxypeptidase.

  1982cited by this paper
• Crystallographic study of the binding of dipeptide inhibitors to thermolysin: implications for the mechanism of catalysis.

  1977cited by this paper
• The structure of thermolysin: an electron density map at 2-3 A resolution.

  1972cited by this paper
• A dipeptidyl carboxypeptidase that converts angiotensin I and inactivates bradykinin.

  1970cited by this paper
• THE PREPARATION AND FUNCTION OF THE HYPERTENSIN-CONVERTING ENZYME

  1956cited by this paper

• A Novel Pathway Implicated in Regulating Cognitive disfunction in a Drosophila Alzheimer’s Disease Model through Acer Inhibition and CG2233 Modulation

  2023cites this paper
• Inhibition of Angiotensin Converting Enzyme by Phytochemicals in Cucurbita pepo L.: In silico Approach

  2022cites this paper
• Enzymes in the time of COVID‐19: An overview about the effects in the human body, enzyme market, and perspectives for new drugs

  2022cites this paper
• Ubiquitin Carboxyl-Terminal Hydrolases and Human Malignancies: The Novel Prognostic and Therapeutic Implications for Head and Neck Cancer

  2021cites this paper
• Targeting UCHL1 Induces Cell Cycle Arrest in High-Risk Multiple Myeloma with t(4;14)

  2021cites this paper
• Deubiquitination of CD36 by UCHL1 promotes foam cell formation

  2020cites this paper
• Novel Therapeutic Approaches Targeting the Renin-Angiotensin System and Associated Peptides in Hypertension and Heart Failure

  2019cites this paper
• Identificación in silico de nuevos compuestos inhibidores de Termolisina derivados de la planta Boldoa purpurascens

  2018cites this paper
• The role of DA1 in organ size control in Arabidopsis thaliana

  2017cites this paper
• Natural Angiotensin Converting Enzyme (ACE) Inhibitors with Antihypertensive Properties

  2017cites this paper
• An approach to identify new antihypertensive agents using Thermolysin as model: In silico study based on QSARINS and docking

  2016cites this paper
• Structure based inhibitor design studies on angiotensin converting enzyme homologues

  2015cites this paper
• Radiolabeled gastrin/CCK analogs in tumor diagnosis: towards higher stability and improved tumor targeting.

  2015cites this paper
• Aphid salivary proteases are capable of degrading sieve-tube proteins.

  2015cites this paper
• Structure–inhibition relationship of phenylethanoid glycosides on angiotensin-converting enzyme using ultra-performance liquid chromatography-tandem quadrupole mass spectrometry

  2015cites this paper
• Production of angiotensin converting enzyme (ACE) inhibitory peptides derived from Thai panga skin

  2014cites this paper
• A virtual screening method for inhibitory peptides of Angiotensin I-converting enzyme.

  2014cites this paper
• Molecular Dynamics Simulation and Molecular Docking Studies of Angiotensin Converting Enzyme with Inhibitor Lisinopril and Amyloid Beta Peptide

  2013cites this paper
• Chapter 77 – Introduction: Metallopeptidases and Their Clans

  2013cites this paper
• Characterization of nicotianamine isolated from soybeans.

  2013cites this paper
• Catalytic mechanism of angiotensin-converting enzyme and effects of the chloride ion.

  2013cites this paper
• Isolation of novel bioactive regions from bovine Achilles tendon collagen having angiotensin I-converting enzyme-inhibitory properties

  2012cites this paper
• Excess α-synuclein worsens disease in mice lacking ubiquitin carboxy-terminal hydrolase L1

  2012cites this paper
• Lipopolysaccharide-induced Overproduction of Nitric Oxide and Overexpression of iNOS and Interleukin-1β Proteins in Zinc-deficient Rats

  2012cites this paper
• Inhibition of Angiotensin-Converting Enzyme Activity by Flavonoids: Structure-Activity Relationship Studies

  2012cites this paper
• Evolution of Genes Involved in Gamete Interaction: Evidence for Positive Selection, Duplications and Losses in Vertebrates

  2012cites this paper
• In Vitro Dissolution Kinetics of Captopril from Microspheres Manufactured by Solvent Evaporation

  2012cites this paper
• Zinc-deficient diet aggravates ventilation-induced lung injury in rats

  2012cites this paper
• Coordination ability of bradykinin with ZnII- and AgI-metal ions – Experimental and theoretical study

  2012cites this paper
• Purification and characterization of ram epididymal angiotensin converting enzyme

  2012cites this paper
• Histone H2A deubiquitinase activity of the Polycomb repressive complex PR-DUB

  2010cites this paper
• Probing the catalytically essential residues of a recombinant dipeptidyl carboxypeptidase from Escherichia coli

  2010cites this paper
• High-resolution crystal structures of Drosophila melanogaster angiotensin-converting enzyme in complex with novel inhibitors and antihypertensive drugs.

  2010cites this paper
• Theoretical and experimental studies of thermolysin inhibition

  2008cites this paper
• Angiotensin-converting enzyme in Spodoptera littoralis: molecular characterization, expression and activity profile during development.

  2008cites this paper
• Conception et synthèse d'inhibiteurs de métalloprotéases et de cibles à ligand acide

  2008cites this paper
• The MEROPS batch BLAST: a tool to detect peptidases and their non-peptidase homologues in a genome.

  2008cites this paper
• Expression, purification, and physicochemical characterization of the N‐terminal active site of human angiotensin‐I converting enzyme

  2007cites this paper
• Role of two active site Glu residues in the molecular action of botulinum neurotoxin endopeptidase.

  2007cites this paper
• Identification and characterisation of the angiotensin converting enzyme-3 (ACE3) gene: a novel mammalian homologue of ACE

  2007cites this paper
• Zinc-metalloproteases in insects: ACE and ECE.

  2004cites this paper
• 84 – Angiotensin-converting enzyme 2

  2004cites this paper
• Introduction: metallopeptidases and their clans

  2004cites this paper
• What’s new in the renin-angiotensin system?

  2004cites this paper
• Structural features of angiotensin-I converting enzyme catalytic sites: conformational studies in solution, homology models and comparison with other zinc metallopeptidases.

  2004influential citation
• Zinc binding in peptide models of angiotensin‐I converting enzyme active sites studied through 1H‐NMR and chemical shift perturbation mapping

  2003cites this paper
• Crystal structure of the human angiotensin-converting enzyme–lisinopril complex

  2003cites this paper
• Do angiotensin-converting enzyme inhibitors directly stimulate the kinin B1 receptor?

  2003cites this paper
• Crystal structure of Drosophila angiotensin I‐converting enzyme bound to captopril and lisinopril 1

  2003cites this paper
• Structure-function discrimination of the N- and C- catalytic domains of human angiotensin-converting enzyme: implications for Cl- activation and peptide hydrolysis mechanisms.

  2003cites this paper
• Angiotensin-converting enzyme-2 (ACE2): comparative modeling of the active site, specificity requirements, and chloride dependence.

  2003cites this paper
• Maintenance of normal blood pressure and renal functions are independent effects of angiotensin-converting enzyme.

  2003cites this paper
• The angiotensin-converting enzyme gene family: genomics and pharmacology.

  2002cites this paper
• The Germinal Isozyme of Angiotensin-converting Enzyme Can Substitute for the Somatic Isozyme in Maintaining Normal Renal Structure and Functions*

  2002cites this paper
• Contribution of Molecular Modeling and Site-directed Mutagenesis to the Identification of Two Structural Residues, Arg-220 and Asp-227, in Aminopeptidase A*

  2002cites this paper
• Novel peptidomimics as angiotensin-converting enzyme inhibitors: a combinatorial approach.

  2002cites this paper
• Crystal structures of leukotriene A4 hydrolase in complex with captopril and two competitive tight‐binding inhibitors

  2002cites this paper
• Identification of glutamic acid 479 as the gluzincin coordinator of zinc in FtsH (HflB).

  2002cites this paper
• Site-directed mutagenesis identifies active-site residues of the light chain of botulinum neurotoxin type A.

  2001cites this paper
• Angiotensin I-converting Enzyme Transition State Stabilization by His1089

  2001cites this paper
• Fluorescence Polarization Studies of Different Forms of Angiotensin-Converting Enzyme

  2001cites this paper
• Isolation and expression of the ecdysteroid-inducible angiotensin-converting enzyme-related gene in wing discs of Bombyx mori.

  2001cites this paper
• A Human Homolog of Angiotensin-converting Enzyme

  2000cites this paper
• Exclusion of Angiotensin I-Converting Enzyme as a Candidate Gene Involved In Exudative Inflammatory Resistance in F344/N Rats

  2000cites this paper
• Molecular and Physiological Aspects of Angiotensin I Converting Enzyme

  1999cites this paper
• Endocrine Control of Sodium Balance

  1999cites this paper
• Delapril versus enalapril in patients with congestive heart failure

  1999cites this paper
• Nitric oxide synthase genes: candidate genes among many others.

  1999cites this paper
• Identification and chromosomal location of two human genes encoding enzymes potentially involved in proteolytic maturation of farnesylated proteins.

  1999cites this paper
• Intragenic deletion in the gene encoding ubiquitin carboxy-terminal hydrolase in gad mice

  1999cites this paper
• Potentiation of Bradykinin Actions by ACE Inhibitors.

  1999cites this paper
• N-domain selectivity of angiotensin I-converting enzyme as assessed by structure-function studies of its highly selective substrate, N-acetyl-seryl-aspartyl-lysyl-proline.

  1999cites this paper
• Zinc Coordination and Substrate Catalysis within the Neuropeptide Processing Enzyme Endopeptidase EC 3.4.24.15

  1999cites this paper
• Kinetic evaluation of beta-neoendorphin hydrolysis by the somatic and testicular isozymes of human angiotensin-converting enzyme.

  1997cites this paper
• Meta-substituted benzofused macrocyclic lactams as zinc metalloprotease inhibitors.

  1997cites this paper
• Cloning and characterisation of angiotensin-converting enzyme from the dipteran species, Haematobia irritans exigua, and its expression in the maturing male reproductive system.

  1996cites this paper
• Affinity of angiotensin I-converting enzyme (ACE) inhibitors for N- and C-binding sites of human ACE is different in heart, lung, arteries, and veins.

  1996cites this paper
• Identification of Glutamate Residues Essential for Catalytic Activity and Zinc Coordination in Aminopeptidase A (*)

  1996cites this paper
• 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.

  1996cites this paper
• A study of chimeras constructed with the two domains of angiotensin I-converting enzyme.

  1996cites this paper
• Peptidyl dipeptidase A: angiotensin I-converting enzyme.

  1995cites this paper
• Recent advances in knowledge of the structure and function of the angiotensin I converting enzyme

  1995cites this paper
• Cloning and Expression of an Evolutionary Conserved Single-domain Angiotensin Converting Enzyme from Drosophila melanogaster(*)

  1995cites this paper
• Isolation and expression of the ecdysteroid-inducible angiotensin-converting enzyme-related gene in wing discs of B mbyx mori

  year unknowncites this paper