Activation of cellular signaling systems is increasingly recognized as a potential modulator of not only the function of plasma membrane proteins, but also their trafficking into and out of the cell membrane. In the heart, Na+ channels are pivotal to both normal electrophysiology and the genesis of life-threatening cardiac arrhythmias. While cardiac Na+ current is modulated by protein kinase(pKA), the nature of this regulation is controversial and the mechanisms unknown. Based on data we have generated, the goal of this research is to test the hypothesis that PKA activation regulates trafficking of cardiac Na+ channels and to investigate the molecular mechanisms. In cells expressing the human cardiac Na+ channel hHl and in rat ventricular myocytes, the effects of pKA activation on N a+ current, plasma membrane channel density, and the cellular distribution of Na+ channels will be determined using electrophysiologic and biochemical methods, immunolocalization, and a fluorescent channel fusion protein to visualize trafficking in living cells. Our preliminary data have implicated the I-II interdomain linker as a region contributing to this effect. To investigate the structural and molecular components required for channel recycling under basal and stimulated states, structure- function hypotheses will be explored for this region as well as the carboxy terminus, which interacts with other proteins (e.g., those with PDZ-domains). Evidence for interacting or adaptor proteins will be sought by both overexpression and binding studies using the channel region(s) involved in the PKA effect. To explore the cellular mechanisms that mediate regulated trafficking of Na+ channels, the role of specific molecular components in both exocytotic and endocytotic pathways will be investigated. The knowledge gained from these studies will improve our understanding of the molecular mechanisms whereby cardiac cells regulate Na+ channel activity and could identify novel strategies to modulate cardiac Na+ currents.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL055665-08
Application #
6750166
Study Section
Cardiovascular and Renal Study Section (CVB)
Program Officer
Wang, Lan-Hsiang
Project Start
1997-04-01
Project End
2006-06-30
Budget Start
2004-07-01
Budget End
2006-06-30
Support Year
8
Fiscal Year
2004
Total Cost
$339,750
Indirect Cost
Name
Vanderbilt University Medical Center
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004413456
City
Nashville
State
TN
Country
United States
Zip Code
37212
Hallaq, Haifa; Wang, Dao W; Kunic, Jennifer D et al. (2012) Activation of protein kinase C alters the intracellular distribution and mobility of cardiac Na+ channels. Am J Physiol Heart Circ Physiol 302:H782-9
Yang, Zhenjiang; Murray, Katherine T (2011) Ionic mechanisms of pacemaker activity in spontaneously contracting atrial HL-1 cells. J Cardiovasc Pharmacol 57:28-36
Wang, Limin; Gill, Rajan; Pedersen, Theresa L et al. (2009) Triglyceride-rich lipoprotein lipolysis releases neutral and oxidized FFAs that induce endothelial cell inflammation. J Lipid Res 50:204-13
Wang, Limin; Sapuri-Butti, Annapoorna R; Aung, Hnin Hnin et al. (2008) Triglyceride-rich lipoprotein lipolysis increases aggregation of endothelial cell membrane microdomains and produces reactive oxygen species. Am J Physiol Heart Circ Physiol 295:H237-44
Yang, Zhenjiang; Browning, Carrie F; Hallaq, Haifa et al. (2008) Four and a half LIM protein 1: a partner for KCNA5 in human atrium. Cardiovasc Res 78:449-57
Hallaq, Haifa; Yang, Zhenjiang; Viswanathan, Prakash C et al. (2006) Quantitation of protein kinase A-mediated trafficking of cardiac sodium channels in living cells. Cardiovasc Res 72:250-61
Zhou, Jingsong; Shin, Hyeon-Gyu; Yi, Jianxun et al. (2002) Phosphorylation and putative ER retention signals are required for protein kinase A-mediated potentiation of cardiac sodium current. Circ Res 91:540-6
Shin, H G; Murray, K T (2001) Conventional protein kinase C isoforms and cross-activation of protein kinase A regulate cardiac Na+ current. FEBS Lett 495:154-8
Shin, H G; Barnett, J V; Chang, P et al. (2000) Molecular heterogeneity of protein kinase C expression in human ventricle. Cardiovasc Res 48:285-99
Zhou, J; Yi, J; Hu, N et al. (2000) Activation of protein kinase A modulates trafficking of the human cardiac sodium channel in Xenopus oocytes. Circ Res 87:33-8