Endothelial Nitric Oxide Synthase Phosphorylation at Threonine 495 and Mitochondrial Reactive Oxygen Species Formation in Response to a High H2O2 ConcentrationGuterbaum T.J.a, b · Braunstein T.H.a, b · Fossum A.c · Holstein-Rathlou N.-H.a, b · Torp-Pedersen C.T.a,b,d · Domínguez H.a,b,e
aThe Danish National Research Foundation Center for Cardiac Arrhythmia and bDepartment of Biomedical Sciences, Faculty of Health Sciences, and cBiotech Research and Innovation Center, FACS Facility, University of Copenhagen, Copenhagen, dCardiology Department P, Research Unit, Gentofte University Hospital, Hellerup, and eCardiology Department S, Research Unit, Herlev University Hospital, Herlev, Denmark
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Background: Hydrogen peroxide (H2O2) is produced in vessels during ischemia/reperfusion and during inflammation, both leading to vascular dysfunction. We investigated cellular pathways involved in endothelial nitric oxide synthase (eNOS) phosphorylation at Threonine 495 (Thr495) in human umbilical vein endothelial cells (HUVECs) exposed to H2O2. Methods: HUVECs were exposed to 400 μ
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- Furchgott RF: The 1996 Albert Lasker Medical Research Awards. The discovery of endothelium-derived relaxing factor and its importance in the identification of nitric oxide. JAMA 1996;276:1186-1188.
- Fleming I, Busse R: NO: the primary EDRF. J Mol Cell Cardiol 1999;31:5-14.
- Napoli C, de Nigris F, Williams-Ignarro S, et al: Nitric oxide and atherosclerosis: an update. Nitric Oxide 2006;15:265-279.
- Cai H, Harrison DG: Endothelial dysfunction in cardiovascular diseases: the role of oxidant stress. Circ Res 2000;87:840-844.
- Pacher P, Beckman JS, Liaudet L: Nitric oxide and peroxynitrite in health and disease. Physiol Rev 2007;87:315-424.
- Srinivasan S, Hatley ME, Bolick DT, et al: Hyperglycaemia-induced superoxide production decreases eNOS expression via AP-1 activation in aortic endothelial cells. Diabetologia 2004;47:1727-1734.
- Forstermann U, Munzel T: Endothelial nitric oxide synthase in vascular disease: from marvel to menace. Circulation 2006;113:1708-1714.
- Hensley K, Maidt ML, Yu Z, et al: Electrochemical analysis of protein nitrotyrosine and dityrosine in the Alzheimer brain indicates region-specific accumulation. J Neurosci 1998;18:8126-8132.
- Tota B, Quintieri AM, Angelone T: The emerging role of nitrite as an endogenous modulator and therapeutic agent of cardiovascular function. Curr Med Chem 2010;17:1915-1925.
- Davidson SM, Duchen MR: Endothelial mitochondria: contributing to vascular function and disease. Circ Res 2007;100:1128-1141.
- Droge W: Free radicals in the physiological control of cell function. Physiol Rev 2002;82:47-95.
- Zhuang S, Kinsey GR, Yan Y, et al: Extracellular signal-regulated kinase activation mediates mitochondrial dysfunction and necrosis induced by hydrogen peroxide in renal proximal tubular cells. J Pharmacol Exp Ther 2008;325:732-740.
- Murphy MP: How mitochondria produce reactive oxygen species. Biochem J 2009;417:1-13.
- Kudin AP, Bimpong-Buta NY, Vielhaber S, et al: Characterization of superoxide-producing sites in isolated brain mitochondria. J Biol Chem 2004;279:4127-4135.
- Teng JC, Kay H, Chen Q, et al: Mechanisms related to the cardioprotective effects of protein kinase C epsilon (PKC epsilon) peptide activator or inhibitor in rat ischemia/reperfusion injury. Naunyn Schmiedebergs Arch Pharmacol 2008;378:1-15.
- Coyle CH, Martinez LJ, Coleman MC, et al: Mechanisms of H2O2-induced oxidative stress in endothelial cells. Free Radic Biol Med 2006;40:2206-2213.
- Shimokawa H, Morikawa K: Hydrogen peroxide is an endothelium-derived hyperpolarizing factor in animals and humans. J Mol Cell Cardiol 2005;39:725-732.
- Sugimoto M, Nakayama M, Goto TM, et al: Rho-kinase phosphorylates eNOS at threonine 495 in endothelial cells. Biochem Biophys Res Commun 2007;361:462-467.
- Fleming I, Busse R: Signal transduction of eNOS activation. Cardiovasc Res 1999;43:532-541.
- Michell BJ, Chen Z, Tiganis T, et al: Coordinated control of endothelial nitric-oxide synthase phosphorylation by protein kinase C and the cAMP-dependent protein kinase. J Biol Chem 2001;276:17625-17628.
- Dimmeler S, Fleming I, Fisslthaler B, et al: Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation. Nature 1999;399:601-605.
- Cai H, Li Z, Davis ME, et al: Akt-dependent phosphorylation of serine 1179 and mitogen-activated protein kinase kinase/extracellular signal-regulated kinase 1/2 cooperatively mediate activation of the endothelial nitric-oxide synthase by hydrogen peroxide. Mol Pharmacol 2003;63:325-331.
- Thomas SR, Chen K, Keaney JF Jr: Hydrogen peroxide activates endothelial nitric-oxide synthase through coordinated phosphorylation and dephosphorylation via a phosphoinositide 3-kinase-dependent signaling pathway. J Biol Chem 2002;277:6017-6024.
- Hu Z, Chen J, Wei Q, et al: Bidirectional actions of hydrogen peroxide on endothelial nitric-oxide synthase phosphorylation and function: co-commitment and interplay of Akt and AMPK. J Biol Chem 2008;283:25256-25263.
- Kobara M, Tatsumi T, Takeda M, et al: The dual effects of nitric oxide synthase inhibitors on ischemia-reperfusion injury in rat hearts. Basic Res Cardiol 2003;98:319-328.
- Rabelo LA, Cortes SF, Alvarez-Leite JI, et al: Endothelium dysfunction in LDL receptor knockout mice: a role for H2O2. Br J Pharmacol 2003;138:1215-1220.
- Yada T, Shimokawa H, Hiramatsu O, et al: Hydrogen peroxide, an endogenous endothelium-derived hyperpolarizing factor, plays an important role in coronary autoregulation in vivo. Circulation 2003;107:1040-1045.
- Lebuffe G, Schumacker PT, Shao ZH, et al: ROS and NO trigger early preconditioning: relationship to mitochondrial KATP channel. Am J Physiol Heart Circ Physiol 2003;284:H299-H308.
- Tang XQ, Chen J, Tang EH, et al: Hydrogen peroxide preconditioning protects PC12 cells against apoptosis induced by oxidative stress. Sheng Li Xue Bao 2005;57:211-216.
- Jugdutt BI: Nitric oxide and cardioprotection during ischemia-reperfusion. Heart Fail Rev 2002;7:391-405.
- Hare JM, Stamler JS: NO/redox disequilibrium in the failing heart and cardiovascular system. J Clin Invest 2005;115:509-517.
- Boyle JJ, Harrington HA, Piper E, et al: Coronary intraplaque hemorrhage evokes a novel atheroprotective macrophage phenotype. Am J Pathol 2009;174:1097-1108.
- Meuwissen M, van der Wal AC, Niessen HW, et al: Colocalisation of intraplaque C reactive protein, complement, oxidised low density lipoprotein, and macrophages in stable and unstable angina and acute myocardial infarction. J Clin Pathol 2006;59:196-201.
- Halliwell B, Clement MV, Long LH: Hydrogen peroxide in the human body. FEBS Lett 2000;486:10-13.
- Liu X, Zweier JL: A real-time electrochemical technique for measurement of cellular hydrogen peroxide generation and consumption: evaluation in human polymorphonuclear leukocytes. Free Radic Biol Med 2001;31:894-901.
- Anzalone R, La Rocca G, Di Stefano A, et al: Role of endothelial cell stress in the pathogenesis of chronic heart failure. Front Biosci 2009;14:2238-2247.
- Azevedo-Martins AK, Lortz S, Lenzen S, et al: Improvement of the mitochondrial antioxidant defense status prevents cytokine-induced nuclear factor-kappaB activation in insulin-producing cells. Diabetes 2003;52:93-101.
- Ali MH, Mungai PT, Schumacker PT: Stretch-induced phosphorylation of focal adhesion kinase in endothelial cells: role of mitochondrial oxidants. Am J Physiol Lung Cell Mol Physiol 2006;291:L38-L45.
- Kelley EE, Khoo NK, Hundley NJ, et al: Hydrogen peroxide is the major oxidant product of xanthine oxidase. Free Radic Biol Med 2010;48:493-498.
- Cortese-Krott MM, Rodriguez-Mateos A, Kuhnle GG, et al: A multilevel analytical approach for detection and visualization of intracellular NO production and nitrosation events using diaminofluoresceins. Free Radic Biol Med 2012;53:2146-2158.
- Kamei K, Haruyama T, Mie M, et al: The construction of endothelial cellular biosensing system for the control of blood pressure drugs. Biosens Bioelectron 2004;19:1121-1124.
- Payne GA, Bohlen HG, Dincer UD, et al: Periadventitial adipose tissue impairs coronary endothelial function via PKC-beta-dependent phosphorylation of nitric oxide synthase. Am J Physiol Heart Circ Physiol 2009;297:H460-H465.
- Jirousek MR, Gillig JR, Gonzalez CM, et al: (S)-13-[(dimethylamino)methyl]-10,11,14,15- tetrahydro-4,9:16, 21-dimetheno-1H, 13H-dibenzo[e,k]pyrrolo[3,4-h][1,4,13]oxadiazacyclohexadecene-1,3(2H)-d ione (LY333531) and related analogues: isozyme selective inhibitors of protein kinase C beta. J Med Chem 1996;39:2664-2671.
- Craig LE, Spelman JP, Strandberg JD, et al: Endothelial cells from diverse tissues exhibit differences in growth and morphology. Microvasc Res 1998;55:65-76.
- Chi JT, Chang HY, Haraldsen G, et al: Endothelial cell diversity revealed by global expression profiling. Proc Natl Acad Sci USA 2003;100:10623-10628.
- Takemoto M, Sun J, Hiroki J, et al: Rho-kinase mediates hypoxia-induced downregulation of endothelial nitric oxide synthase. Circulation 2002;106:57-62.
- Cale JM, Bird IM: Inhibition of MEK/ERK1/2 signalling alters endothelial nitric oxide synthase activity in an agonist-dependent manner. Biochem J 2006;398:279-288.
- Hattori Y, Matsumura M, Kasai K: Vascular smooth muscle cell activation by C-reactive protein. Cardiovasc Res 2003;58:186-195.
- Leikert JF, Rathel TR, Wohlfart P, et al: Red wine polyphenols enhance endothelial nitric oxide synthase expression and subsequent nitric oxide release from endothelial cells. Circulation 2002;106:1614-1617.
- Li H, Junk P, Huwiler A, et al: Dual effect of ceramide on human endothelial cells: induction of oxidative stress and transcriptional upregulation of endothelial nitric oxide synthase. Circulation 2002;106:2250-2256.
- Yoshizumi M, Perrella MA, Burnett JC Jr, et al: Tumor necrosis factor downregulates an endothelial nitric oxide synthase mRNA by shortening its half-life. Circ Res 1993;73:205-209.
- Pirkmajer S, Chibalin AV: Serum starvation: caveat emptor. Am J Physiol Cell Physiol 2011;301:C272-C279.
- Holland JA, Meyer JW, Chang MM, et al: Thrombin stimulated reactive oxygen species production in cultured human endothelial cells. Endothelium 1998;6:113-121.
- Davies SP, Reddy H, Caivano M, et al: Specificity and mechanism of action of some commonly used protein kinase inhibitors. Biochem J 2000;351:95-105.
- Wang X, Martindale JL, Liu Y, et al: The cellular response to oxidative stress: influences of mitogen-activated protein kinase signalling pathways on cell survival. Biochem J 1998;333:291-300.
- Lee WC, Choi CH, Cha SH, et al: Role of ERK in hydrogen peroxide-induced cell death of human glioma cells. Neurochem Res 2005;30:263-270.
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