Angiotensin Converting Enzyme (ACE) and ACE2 Bind Integrins and ACE2 Regulates Integrin Signalling

The angiotensin converting enzymes (ACEs) are the key catalytic components of the renin-angiotensin system, mediating precise regulation of blood pressure by counterbalancing the effects of each other. Inhibition of ACE has been shown to improve pathology in cardiovascular disease, whilst ACE2 is cardioprotective in the failing heart. However, the mechanisms by which ACE2 mediates its cardioprotective functions have yet to be fully elucidated. Here we demonstrate that both ACE and ACE2 bind integrin subunits, in an RGD-independent manner, and that they can act as cell adhesion substrates. We show that cellular expression of ACE2 enhanced cell adhesion. Furthermore, we present evidence that soluble ACE2 (sACE2) is capable of suppressing integrin signalling mediated by FAK. In addition, sACE2 increases the expression of Akt, thereby lowering the proportion of the signalling molecule phosphorylated Akt. These results suggest that ACE2 plays a role in cell-cell interactions, possibly acting to fine-tune integrin signalling. Hence the expression and cleavage of ACE2 at the plasma membrane may influence cell-extracellular matrix interactions and the signalling that mediates cell survival and proliferation. As such, ectodomain shedding of ACE2 may play a role in the process of pathological cardiac remodelling.

[1]  E. Sturrock,et al.  Epitope mapping of mAbs to denatured human testicular ACE (CD143). , 2008, Tissue antigens.

[2]  Qingbo Xu,et al.  Different Effects of Angiotensin II and Angiotensin-(1-7) on Vascular Smooth Muscle Cell Proliferation and Migration , 2010, PloS one.

[3]  E. Jiménez,et al.  Angiotensin II induces MMP 2 activity via FAK/JNK pathway in human endothelial cells. , 2009, Biochemical and biophysical research communications.

[4]  Johan Verjans,et al.  Myocardial remodeling after infarction: the role of myofibroblasts , 2010, Nature Reviews Cardiology.

[5]  Junichi Takagi,et al.  Structural basis for ligand recognition by integrins. , 2007, Current opinion in cell biology.

[6]  Wei Li,et al.  Effects of Early Angiotensin-Converting Enzyme Inhibition on Cardiac Gene Expression After Acute Myocardial Infarction , 2001, Circulation.

[7]  Shokei Kim,et al.  Beneficial effects of combination of ACE inhibitor and angiotensin II type 1 receptor blocker on cardiac remodeling in rat myocardial infarction. , 2003, Cardiovascular research.

[8]  B. Greenberg,et al.  Angiotensin-(1-7) binds to specific receptors on cardiac fibroblasts to initiate antifibrotic and antitrophic effects. , 2005, American journal of physiology. Heart and circulatory physiology.

[9]  J. Penninger,et al.  Loss of angiotensin-converting enzyme-2 (Ace2) accelerates diabetic kidney injury. , 2007, The American journal of pathology.

[10]  T. Sasazuki,et al.  Modulation of TNF-α-converting enzyme by the spike protein of SARS-CoV and ACE2 induces TNF-α production and facilitates viral entry , 2008, Proceedings of the National Academy of Sciences.

[11]  D. Selkoe,et al.  Secreted APP regulates the function of full-length APP in neurite outgrowth through interaction with integrin beta1 , 2008, Neural Development.

[12]  P. K. Smith,et al.  Measurement of protein using bicinchoninic acid. , 1985, Analytical biochemistry.

[13]  M. Raizada,et al.  ACE2 overexpression inhibits hypoxia-induced collagen production by cardiac fibroblasts. , 2007, Clinical science.

[14]  M. Goddard,et al.  ACE2 gene expression is up-regulated in the human failing heart , 2004, BMC medicine.

[15]  M. Bristow,et al.  Angiotensin-(1-7) Formation in the Intact Human Heart: In Vivo Dependence on Angiotensin II as Substrate , 2003, Circulation.

[16]  A. Turner,et al.  Not just angiotensinases: new roles for the angiotensin-converting enzymes , 2009, Cellular and Molecular Life Sciences.

[17]  S. Hanks,et al.  Focal adhesion protein-tyrosine kinase phosphorylated in response to cell attachment to fibronectin. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[18]  Nigel M. Hooper,et al.  Calmodulin interacts with angiotensin‐converting enzyme‐2 (ACE2) and inhibits shedding of its ectodomain , 2007, FEBS Letters.

[19]  C. Ferrario,et al.  Angiotensin-(1-7) inhibits growth of cardiac myocytes through activation of the mas receptor. , 2005, American journal of physiology. Heart and circulatory physiology.

[20]  T. Walther,et al.  Angiotensin(1-7) Blunts Hypertensive Cardiac Remodeling by a Direct Effect on the Heart , 2008, Circulation research.

[21]  P. Hillemanns,et al.  A beta 1-integrin receptor for fibronectin in human kidney glomeruli. , 1989, The American journal of pathology.

[22]  S. Ball,et al.  Simvastatin reduces human atrial myofibroblast proliferation independently of cholesterol lowering via inhibition of RhoA. , 2004, Cardiovascular research.

[23]  D. Turk,et al.  The refined 1.9‐Å X‐ray crystal structure of d‐Phe‐Pro‐Arg chloromethylketone‐inhibited human α‐thrombin: Structure analysis, overall structure, electrostatic properties, detailed active‐site geometry, and structure‐function relationships , 1992, Protein science : a publication of the Protein Society.

[24]  Duncan J. Stewart,et al.  Infarction : Regulation by Cytokines 3 in the Heart After Myocardial β 1 and β Temporal Response and Localization of Integrins , 2003 .

[25]  K. Griendling,et al.  NADPH oxidases and angiotensin II receptor signaling , 2009, Molecular and Cellular Endocrinology.

[26]  Nigel M. Hooper,et al.  A Human Homolog of Angiotensin-converting Enzyme , 2000, The Journal of Biological Chemistry.

[27]  Thomas Walther,et al.  Angiotensin-(1–7) is an endogenous ligand for the G protein-coupled receptor Mas , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[28]  F. Beau,et al.  Development of potent and selective phosphinic peptide inhibitors of angiotensin-converting enzyme 2. , 2008, Journal of medicinal chemistry.

[29]  Q. Lin,et al.  Interaction of ACE2 and integrin beta1 in failing human heart. , 2004, Biochimica et biophysica acta.

[30]  M. Cattaneo,et al.  Anti-migratory and Anti-invasive Effect of Somatostatin in Human Neuroblastoma Cells , 2003, Journal of Biological Chemistry.

[31]  Shokei Kim,et al.  Angiotensin converting enzyme inhibitor prevents left ventricular remodelling after myocardial infarction in angiotensin II type 1 receptor knockout mice , 2005, Heart.

[32]  F. Rosenow,et al.  Antimetastatic Integrin as Inhibitors of Snake Venoms , 2008 .

[33]  M. Raizada,et al.  Cardiac Overexpression of Angiotensin Converting Enzyme 2 Protects the Heart From Ischemia-Induced Pathophysiology , 2008, Hypertension.

[34]  F. Rosenow,et al.  Integrins as antimetastatic targets of RGD-independent snake venom components in liver metastasis [corrected]. , 2008, Neoplasia.

[35]  J. Grobe,et al.  Chronic angiotensin-(1-7) prevents cardiac fibrosis in DOCA-salt model of hypertension. , 2006, American journal of physiology. Heart and circulatory physiology.

[36]  A. Klein,et al.  Soluble angiotensin-converting enzyme 2 in human heart failure: relation with myocardial function and clinical outcomes. , 2009, Journal of cardiac failure.

[37]  T. Ogihara,et al.  Deletion of Angiotensin-Converting Enzyme 2 Accelerates Pressure Overload–Induced Cardiac Dysfunction by Increasing Local Angiotensin II , 2006, Hypertension.

[38]  Eric O Long.  ICAM-1: Getting a Grip on Leukocyte Adhesion , 2011, The Journal of Immunology.

[39]  N. Hooper,et al.  Tumor Necrosis Factor-α Convertase (ADAM17) Mediates Regulated Ectodomain Shedding of the Severe-acute Respiratory Syndrome-Coronavirus (SARS-CoV) Receptor, Angiotensin-converting Enzyme-2 (ACE2) , 2005, Journal of Biological Chemistry.

[40]  J. McDonald,et al.  Expression of Fibronectin, the Integrin α5, and α-Smooth Muscle Actin in Heart and Lung Development , 1992 .

[41]  Robert Kleta,et al.  Tissue-Specific Amino Acid Transporter Partners ACE2 and Collectrin Differentially Interact With Hartnup Mutations , 2008, Gastroenterology.

[42]  Jodie L. Guy,et al.  Residues affecting the chloride regulation and substrate selectivity of the angiotensin‐converting enzymes (ACE and ACE2) identified by site‐directed mutagenesis , 2008, The FEBS journal.

[43]  Jodie L. Guy,et al.  Functional angiotensin‐converting enzyme 2 is expressed in human cardiac myofibroblasts , 2008, Experimental physiology.

[44]  N. Sampson,et al.  Mediation of sperm-egg fusion: evidence that mouse egg alpha6beta1 integrin is the receptor for sperm fertilinbeta. , 1999, Chemistry & biology.

[45]  J. Penninger,et al.  Loss of Angiotensin-Converting Enzyme 2 Accelerates Maladaptive Left Ventricular Remodeling in Response to Myocardial Infarction , 2009, Circulation. Heart failure.

[46]  P. Zigrino,et al.  The Disintegrin-like and Cysteine-rich domains of ADAM-9 Mediate Interactions between Melanoma Cells and Fibroblasts* , 2010, The Journal of Biological Chemistry.

[47]  J. Esko,et al.  Thrombin adhesive properties: induction by plasmin and heparan sulfate , 1993, The Journal of cell biology.

[48]  Q. Lin,et al.  Interaction of ACE2 and integrin β1 in failing human heart , 2004 .

[49]  I. Balyasnikova,et al.  Monoclonal antibodies to denatured human ACE (CD 143), broad species specificity, reactivity on paraffin sections, and detection of subtle conformational changes in the C-terminal domain of ACE. , 2003, Tissue antigens.

[50]  M. Lampugnani,et al.  An Arg-Gly-Asp sequence within thrombin promotes endothelial cell adhesion , 1991, The Journal of cell biology.

[51]  N. Turner,et al.  Cardiac fibroblasts: at the heart of myocardial remodeling. , 2009, Pharmacology & therapeutics.

[52]  J. Mocco,et al.  Lentivirus‐mediated overexpression of angiotensin‐(1–7) attenuated ischaemia‐induced cardiac pathophysiology , 2011, Experimental physiology.

[53]  A. Turner,et al.  Angiotensin-Converting Enzyme 2: The First Decade , 2011, International journal of hypertension.

[54]  R. Passier,et al.  Expression and localization of renin and angiotensinogen in rat heart after myocardial infarction. , 1996, The American journal of physiology.

[55]  N. Sampson,et al.  Mediation of sperm-egg fusion: evidence that mouse egg α6β1 integrin is the receptor for sperm fertilinβ , 1999 .

[56]  G. Díaz-Araya,et al.  Enalapril Attenuates Downregulation of Angiotensin-Converting Enzyme 2 in the Late Phase of Ventricular Dysfunction in Myocardial Infarcted Rat , 2006, Hypertension.

[57]  Merlin C. Thomas,et al.  Circulating ACE2 activity is increased in patients with type 1 diabetes and vascular complications , 2012, Journal of hypertension.

[58]  Zhidong Wang,et al.  Angiotensin II Induces Inflammatory Response Partly Via Toll-Like Receptor 4-Dependent Signaling Pathway in Vascular Smooth Muscle Cells , 2009, Cellular Physiology and Biochemistry.

[59]  S. Takeda,et al.  Crystal structures of VAP1 reveal ADAMs' MDC domain architecture and its unique C‐shaped scaffold , 2006, The EMBO journal.

[60]  S. Ball,et al.  Chronic beta2-adrenergic receptor stimulation increases proliferation of human cardiac fibroblasts via an autocrine mechanism. , 2003, Cardiovascular research.

[61]  Mahipal Singh,et al.  Deficiency of β1 integrins results in increased myocardial dysfunction after myocardial infarction , 2006, Heart.

[62]  Saurabh Menon,et al.  ACE2 X-Ray Structures Reveal a Large Hinge-bending Motion Important for Inhibitor Binding and Catalysis , 2004, Journal of Biological Chemistry.

[63]  P. Macdonald,et al.  Myocardial infarction increases ACE2 expression in rat and humans. , 2005, European heart journal.