title: G-Protein-Coupled Receptor MrgD Is a Receptor for Angiotensin-(1-7) Involving Adenylyl Cyclase, cAMP, and Phosphokinase A.
abstract: Angiotensin (Ang)-(1-7) has cardiovascular protective effects and is the opponent of the often detrimental Ang II within the renin-angiotensin system. Although it is well accepted that the G-protein-coupled receptor Mas is a receptor for the heptapeptide, the lack in knowing initial signaling molecules stimulated by Ang-(1-7) prevented definitive characterization of ligand/receptor pharmacology as well as identification of further hypothesized receptors for the heptapeptide. The study aimed to identify a second messenger stimulated by Ang-(1-7) allowing confirmation as well as discovery of the heptapeptide's receptors. Ang-(1-7) elevates cAMP concentration in primary cells, such as endothelial or mesangial cells. Using cAMP as readout in receptor-transfected human embryonic kidney (HEK293) cells, we provided pharmacological proof that Mas is a functional receptor for Ang-(1-7). Moreover, we identified the G-protein-coupled receptor MrgD as a second receptor for Ang-(1-7). Consequently, the heptapeptide failed to increase cAMP concentration in primary mesangial cells with genetic deficiency in both Mas and MrgD Mice deficient in MrgD showed an impaired hemodynamic response after Ang-(1-7) administration. Furthermore, we excluded the Ang II type 2 receptor as a receptor for the heptapeptide but discovered that the Ang II type 2 blocker PD123319 can also block Mas and MrgD receptors. Our results lead to an expansion and partial revision of the renin-angiotensin system, by identifying a second receptor for Ang-(1-7), by excluding Ang II type 2 as a receptor for the heptapeptide, and by enforcing the revisit of such publications which concluded Ang II type 2 function by only using PD123319.
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Anja
authors.initials: A
authors.lastname: Tetzner
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Kinga
authors.initials: K
authors.lastname: Gebolys
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Christian
authors.initials: C
authors.lastname: Meinert
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Sabine
authors.initials: S
authors.lastname: Klein
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Anja
authors.initials: A
authors.lastname: Uhlich
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Jonel
authors.initials: J
authors.lastname: Trebicka
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.).
authors.forename: Óscar
authors.initials: Ó
authors.lastname: Villacañas
authors.affiliation: From the Department of Pharmacology and Therapeutics, School of Medicine, School of Pharmacy, University College Cork (UCC), Cork, Ireland (A.T., K.G., C.M., A.U., T.W.); Departments Obstetrics (A.T., T.W.) and Pediatric Surgery (A.T., T.W.), University of Leipzig, Leipzig, Germany; Institute of Experimental and Clinical Pharmacology and Toxicology, Medical Faculty Mannheim, University Heidelberg, Mannheim, Germany (C.M.); Department of Internal Medicine I, University of Bonn, Bonn, Germany (S.K., J.T.); and Computational Chemistry Department, Intelligent Pharma S.L., Barcelona, Spain (Ó.V.). t.walther@ucc.ie.
authors.forename: Thomas
authors.initials: T
authors.lastname: Walther
mesh_terms: D000262:Adenylyl Cyclases / Q000187:drug effects / Q000378:metabolism*; D000803:Angiotensin I / Q000494:pharmacology*; D000818:Animals; D015153:Blotting, Western; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D005260:Female; D057809:HEK293 Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D006973:Hypertension / Q000188:drug therapy / Q000201:enzymology*; D008297:Male; D050527:Mesangial Cells / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010446:Peptide Fragments / Q000494:pharmacology*; D010770:Phosphotransferases / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000378:metabolism; D012084:Renin-Angiotensin System / Q000187:drug effects; D012680:Sensitivity and Specificity
chemical_list: D010446:Peptide Fragments; D043562:Receptors, G-Protein-Coupled; D000803:Angiotensin I; D010770:Phosphotransferases; D017868:Cyclic AMP-Dependent Protein Kinases; D000262:Adenylyl Cyclases; C118790:angiotensin I (1-7)
keywords: G-protein–coupled receptor; angiotensin; mesangial cells; mice; renin–angiotensin system