title: Angiotensin-Converting Enzyme 2 Metabolizes and Partially Inactivates Pyr-Apelin-13 and Apelin-17: Physiological Effects in the Cardiovascular System.
abstract: Apelin peptides mediate beneficial effects on the cardiovascular system and are being targeted as potential new drugs. However, apelin peptides have extremely short biological half-lives, and improved understanding of apelin peptide metabolism may lead to the discovery of biologically stable analogues with therapeutic potential. We examined the ability of angiotensin-converting enzyme 2 (ACE2) to cleave and inactivate pyr-apelin 13 and apelin 17, the dominant apelin peptides. Computer-assisted modeling shows a conserved binding of pyr-apelin 13 and apelin 17 to the ACE2 catalytic site. In ACE2 knockout mice, hypotensive action of pyr-apelin 13 and apelin 17 was potentiated, with a corresponding greater elevation in plasma apelin levels. Similarly, pharmacological inhibition of ACE2 potentiated the vasodepressor action of apelin peptides. Biochemical analysis confirmed that recombinant human ACE2 can cleave pyr-apelin 13 and apelin 17 efficiently, and apelin peptides are degraded slower in ACE2-deficient plasma. The biological relevance of ACE2-mediated proteolytic processing of apelin peptides was further supported by the reduced potency of pyr-apelin 12 and apelin 16 on the activation of signaling pathways and nitric oxide production from endothelial cells. Importantly, although pyr-apelin 13 and apelin 17 rescued contractile function in a myocardial ischemia-reperfusion model, ACE2 cleavage products, pyr-apelin 12 and 16, were devoid of these cardioprotective effects. We designed and synthesized active apelin analogues that were resistant to ACE2-mediated degradation, thereby confirming that stable apelin analogues can be designed as potential drugs. We conclude that ACE2 represents a major negative regulator of apelin action in the vasculature and heart.
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Wang
authors.initials: W
authors.lastname: Wang
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Shaun M K
authors.initials: SM
authors.lastname: McKinnie
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Maikel
authors.initials: M
authors.lastname: Farhan
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Manish
authors.initials: M
authors.lastname: Paul
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Tyler
authors.initials: T
authors.lastname: McDonald
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Brent
authors.initials: B
authors.lastname: McLean
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Catherine
authors.initials: C
authors.lastname: Llorens-Cortes
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Saugata
authors.initials: S
authors.lastname: Hazra
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: Allan G
authors.initials: AG
authors.lastname: Murray
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.).
authors.forename: John C
authors.initials: JC
authors.lastname: Vederas
authors.affiliation: From the Division of Cardiology, Department of Medicine (W.W., B.M., G.Y.O.), Mazankowski Alberta Heart Institute (W.W., B.M., G.Y.O.), Department of Chemistry, Faculty of Science (S.M.K.M., T.M., J.C.V.), and Division of Nephrology, Department of Medicine (M.F., A.G.M.), University of Alberta, Edmonton, AB, Canada; Department of Zoology, RBC College, West Bengal State University, West Bengal, India (M.P.); INSERM, Laboratory of Central Neuropeptides and Regulations of Water Balance and Cardiovascular Functions, Center for Interdisciplinary Research in Biology, College de France, Paris, France (C.L.-C.); Department of Biotechnology, Indian Institute of Technology, Roorkee, India (S.H.). gavin.oudit@ualberta.ca.
authors.forename: Gavin Y
authors.initials: GY
authors.lastname: Oudit
mesh_terms: D054392:Adipokines / Q000378:metabolism*; D000085962:Angiotensin-Converting Enzyme 2; D000818:Animals; D000073861:Apelin; D002316:Cardiotonic Agents / Q000378:metabolism / Q000494:pharmacology; D002319:Cardiovascular System / Q000187:drug effects / Q000378:metabolism / Q000503:physiopathology; D003198:Computer Simulation; D055808:Drug Discovery; D006207:Half-Life; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism* / Q000494:pharmacology; D051379:Mice; D015428:Myocardial Reperfusion Injury / Q000378:metabolism / Q000517:prevention & control; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D065840:Protective Factors; D011485:Protein Binding / Q000502:physiology; D014661:Vasoconstriction / Q000187:drug effects / Q000502:physiology
chemical_list: D054392:Adipokines; D000073861:Apelin; C402826:Apln protein, mouse; D002316:Cardiotonic Agents; D036341:Intercellular Signaling Peptides and Proteins; C586143:apelin 13, Pyr(1)-; C000621386:apelin 17 peptide, human; D007703:Peptidyl-Dipeptidase A; C000705307:ACE2 protein, human; C000705328:Ace2 protein, mouse; D000085962:Angiotensin-Converting Enzyme 2
keywords: angiotensin-converting enzyme 2; apelin 17; blood pressure; computer-based model; enzyme kinetics; ischemia reperfusion injury; pyr-apelin 13