Accumulating evidence indicates that nitric oxide (NO), through S-nitrosylation of Cys residues within multiple, functionally interrelated signaling elements, regulates agonist-induced desensitization and internalization of B- adrenergic receptors (B-ARs). We have demonstrated that NO synthases and the endogenous S-nitrosothiol (SNO), S-nitrosoglutathione, preserve cardiac contractility and prevent down-regulation of B-ARs during maintained agonist stimulation. These effects of NO/SNO on B-AR signaling and trafficking, which can be recapitulated in cellular systems, appear to be mediated, in significant part, by S-nitrosylation of the G protein-coupled receptor (GPCR) kinase, GRK2. Additional components of the B-AR system are also regulated by S-nitrosylation, which suggests a broad functional role for NO/SNO, exerted through targeted S-nitrosylation. In particular, we have recently identified the B-arrestins (Barr1 and Barr2) as targets of B-AR-coupled S-nitrosylation by NO synthases (eNOS and nNOS). S-nitrosylation of Barr2 by eNOS at a single critical site (Cys 410) regulates its protein-protein interactions with clathrin and adapter protein-2 (AP-2) in vitro and in vivo, thereby promoting agonist-mediated B{2}-AR internalization. However, differential regulation of Barr1 and Barr2 by S-nitrosylation and the consequences of these modifications for cardiac function, have not been explored. Our central hypothesis is that S-nitrosylation of the B-arrestins will provide a basis for control by NO of B-AR trafficking and signaling, with important ramifications in healthy and failing hearts. Moreover, we predict that the regulation of Barr1- and Barr2-specific interactomes by S-nitrosylation will provide a principal mechanism through which NO exerts its regulatory influence. We will carry out the following specific aims:
Specific Aim 1. Elucidate the sites of agonist-dependent S-nitrosylation of the B-arrestins by eNOS and nNOS in cells and tissues.
Specific Aim 2. Elucidate the consequences of B-arrestin S-nitrosylation for B-AR internalization and desensitization.
Specific Aim 3. Assess the consequences of B-arrestin S-nitrosylation for p-AR-dependent signaling.
Specific Aim 4. Assess the functional roles of B-arrestin S-nitrosylation in the intact heart. Collectively, these studies should provide fundamental and novel insights into B-AR regulation by NO in both healthy and failing hearts and may open a new area of research.

National Institute of Health (NIH)
National Heart, Lung, and Blood Institute (NHLBI)
Research Program Projects (P01)
Project #
Application #
Study Section
Heart, Lung, and Blood Program Project Review Committee (HLBP)
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Duke University
United States
Zip Code
Abraham, Dennis M; Davis 3rd, Robert T; Warren, Chad M et al. (2016) β-Arrestin mediates the Frank-Starling mechanism of cardiac contractility. Proc Natl Acad Sci U S A 113:14426-14431
Hullmann, Jonathan; Traynham, Christopher J; Coleman, Ryan C et al. (2016) The expanding GRK interactome: Implications in cardiovascular disease and potential for therapeutic development. Pharmacol Res 110:52-64
Zhang, Rongli; Hess, Douglas T; Reynolds, James D et al. (2016) Hemoglobin S-nitrosylation plays an essential role in cardioprotection. J Clin Invest 126:4654-4658
Woodall, Meryl C; Woodall, Benjamin P; Gao, Erhe et al. (2016) Cardiac Fibroblast GRK2 Deletion Enhances Contractility and Remodeling Following Ischemia/Reperfusion Injury. Circ Res 119:1116-1127
Feldman, Arthur M; Gordon, Jennifer; Wang, JuFang et al. (2016) BAG3 regulates contractility and Ca(2+) homeostasis in adult mouse ventricular myocytes. J Mol Cell Cardiol 92:10-20
Adachi, Naoko; Hess, Douglas T; McLaughlin, Precious et al. (2016) S-Palmitoylation of a Novel Site in the β2-Adrenergic Receptor Associated with a Novel Intracellular Itinerary. J Biol Chem 291:20232-46
Hodavance, Sima Y; Gareri, Clarice; Torok, Rachel D et al. (2016) G Protein-coupled Receptor Biased Agonism. J Cardiovasc Pharmacol 67:193-202
Waldschmidt, Helen V; Homan, Kristoff T; Cruz-Rodríguez, Osvaldo et al. (2016) Structure-Based Design, Synthesis, and Biological Evaluation of Highly Selective and Potent G Protein-Coupled Receptor Kinase 2 Inhibitors. J Med Chem 59:3793-807
Carr 3rd, Richard; Schilling, Justin; Song, Jianliang et al. (2016) β-arrestin-biased signaling through the β2-adrenergic receptor promotes cardiomyocyte contraction. Proc Natl Acad Sci U S A 113:E4107-16
Watson, Lewis J; Alexander, Kevin M; Mohan, Maradumane L et al. (2016) Phosphorylation of Src by phosphoinositide 3-kinase regulates beta-adrenergic receptor-mediated EGFR transactivation. Cell Signal 28:1580-92

Showing the most recent 10 out of 147 publications