zed under Specific Aims 1-4, stem from our initial investigations of enzymes that metabolize peptides such as bradykinin (BK), angiotensins (Ang) and others. Inhibitors of Ang I converting enzyme (kininase II; ACE) are currently used to treat millions of patients for hypertension, and it is widely assumed that their therapeutic action is due to blocking the activation of a hypertensive peptide (Ang II) while inhibiting the inactivation of the hypotensive one (BK). Our research indicates that ACE inhibitors have other important actions as well. They potentiate BK and its peptidase-resistant analogue agonists of the B2 receptor and induce a form of an enzyme-receptor crosstalk. We shall use cultured cells with either transfected or constitutively expressed enzymes and receptors and study, with various techniques, the heterodimer formation between human ACE and the B2 receptor and how ACE inhibitors affect this complex by enhancing BK effects (indirect activation) on the receptor. This leads to an increased release of vascular mediators such as NO and arachidonic acid (prostaglandins;
Specific Aim 1) -- ACE inhibitors also directly activate the B1 receptor of desArg BK and desArg-Lysl-BK (Specific Aims 2 and 3) by combining with a Znbinding pentameric sequence on an extracellular domain of the receptor, in the absence of ACE and peptide agonists. Activation of the B1 receptor (Specific Aims 2 and 3) leads to a prolonged release of NO and possibly other cardiovascular mediators from cells. Their prolonged action on the B1 receptor may affect collagen synthesis. We will investigate the initiation of different transduction pathways by the different modes of activation of the B2 (Specific Aim 1) and B1 receptors using calcium channel blockers and protein kinase inhibitors. We shall continue our study of peptidases and learn more about the structure of human kininase I, which releases the peptide ligands of B1 receptors by cleaving Arg from BK and Lys-Bk.
We aim to crystallize the active subunit of kininase I (carboxypeptidase N) (Specific Aim 4). To obtain mg quantities, we will use an improved fermentation techniques with Pichia pastoris cells. Results of our experiments will help to understand how widely used ACE inhibitors interact with receptors and consequently influence vascular reactivity.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL036473-20
Application #
6892382
Study Section
Cardiovascular and Renal Study Section (CVB)
Program Officer
Rabadan-Diehl, Cristina
Project Start
1993-09-30
Project End
2007-06-30
Budget Start
2005-07-01
Budget End
2006-06-30
Support Year
20
Fiscal Year
2005
Total Cost
$389,675
Indirect Cost
Name
University of Illinois at Chicago
Department
Pharmacology
Type
Schools of Medicine
DUNS #
098987217
City
Chicago
State
IL
Country
United States
Zip Code
60612
Huan, Tianxiao; Joehanes, Roby; Schurmann, Claudia et al. (2016) A whole-blood transcriptome meta-analysis identifies gene expression signatures of cigarette smoking. Hum Mol Genet 25:4611-4623
Lowry, Jessica L; Brovkovych, Viktor; Zhang, Yongkang et al. (2013) Endothelial nitric-oxide synthase activation generates an inducible nitric-oxide synthase-like output of nitric oxide in inflamed endothelium. J Biol Chem 288:4174-93
Zhang, Xianming; Lowry, Jessica L; Brovkovych, Viktor et al. (2012) Characterization of dual agonists for kinin B1 and B2 receptors and their biased activation of B2 receptors. Cell Signal 24:1619-31
Brovkovych, Viktor; Zhang, Yongkang; Brovkovych, Svitlana et al. (2011) A novel pathway for receptor-mediated post-translational activation of inducible nitric oxide synthase. J Cell Mol Med 15:258-69
Erdös, Ervin G; Tan, Fulong; Skidgel, Randal A (2010) Angiotensin I-converting enzyme inhibitors are allosteric enhancers of kinin B1 and B2 receptor function. Hypertension 55:214-20
Skidgel, Randal A; Erdos, Ervin G (2007) Structure and function of human plasma carboxypeptidase N, the anaphylatoxin inactivator. Int Immunopharmacol 7:1888-99
Keil, Cora; Maskos, Klaus; Than, Manuel et al. (2007) Crystal structure of the human carboxypeptidase N (kininase I) catalytic domain. J Mol Biol 366:504-16
Chen, Zhenlong; Deddish, Peter A; Minshall, Richard D et al. (2006) Human ACE and bradykinin B2 receptors form a complex at the plasma membrane. FASEB J 20:2261-70
Stanisavljevic, Sinisa; Ignjatovic, Tatjana; Deddish, Peter A et al. (2006) Angiotensin I-converting enzyme inhibitors block protein kinase C epsilon by activating bradykinin B1 receptors in human endothelial cells. J Pharmacol Exp Ther 316:1153-8
Hecquet, Claudie; Biyashev, Dauren; Tan, Fulong et al. (2006) Positive cooperativity between the thrombin and bradykinin B2 receptors enhances arachidonic acid release. Am J Physiol Heart Circ Physiol 290:H948-58

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