AMP-Activated Protein Kinase Functionally Phosphorylates Endothelial Nitric Oxide Synthase Ser633

2009/01/09 by Zhen Chen, I‐Chen Peng, I-Chen Peng +12 · 27 citations
Biochemistry, Genetics and Molecular Biology · Medicine · #Metabolism, Diabetes, and Cancer #Nitric Oxide and Endothelin Effects #Cardiovascular Function and Risk Factors

paper · pdf · doi:10.1161/circresaha.108.187567

Abstract

Endothelial nitric oxide synthase (eNOS) plays a central role in maintaining cardiovascular homeostasis by controlling NO bioavailability. The activity of eNOS in vascular endothelial cells (ECs) largely depends on posttranslational modifications, including phosphorylation. Because the activity of AMP-activated protein kinase (AMPK) in ECs can be increased by multiple cardiovascular events, we studied the phosphorylation of eNOS Ser633 by AMPK and examined its functional relevance in the mouse models. Shear stress, atorvastatin, and adiponectin all increased AMPK Thr172 and eNOS Ser633 phosphorylations, which were abolished if AMPK was pharmacologically inhibited or genetically ablated. The constitutively active form of AMPK or an AMPK agonist caused a sustained Ser633 phosphorylation. Expression of gain-/loss-of-function eNOS mutants revealed that Ser633 phosphorylation is important for NO production. The aorta of AMPKalpha2(-/-) mice showed attenuated atorvastatin-induced eNOS phosphorylation. Nano-liquid chromatography/tandem mass spectrometry (LC/MS/MS) confirmed that eNOS Ser633 was able to compete with Ser1177 or acetyl-coenzyme A carboxylase Ser79 for AMPKalpha phosphorylation. Nano-LC/MS/MS confirmed that eNOS purified from AICAR-treated ECs was phosphorylated at both Ser633 and Ser1177. Our results indicate that AMPK phosphorylation of eNOS Ser633 is a functional signaling event for NO bioavailability in ECs.

Cited by

• AMPK-mediated regulation of transcription in skeletal muscle
• Extension of lifespan by epicatechin, halofuginone and mitoglitazone in male but not female genetically heterogeneous mice
• Effects of AMP-activated protein kinase in cerebral ischemia. [europepmc]
• Cellular signaling and NO production. [europepmc]
• Activation of AMP-activated protein kinase by vascular endothelial growth factor mediates endothelial angiogenesis independently of nitric-oxide synthase. [europepmc]
• AMPKalpha2 deletion causes aberrant expression and activation of NAD(P)H oxidase and consequent endothelial dysfunction in vivo: role of 26S proteasomes. [europepmc]
• AMPK inhibition in health and disease. [europepmc]
• AMPK in cardiovascular health and disease. [europepmc]
• Effects of metformin in experimental stroke. [europepmc]
• AMP-activated protein kinase and metabolic control. [europepmc]
• Acute exposure to low glucose rapidly induces endothelial dysfunction and mitochondrial oxidative stress: role for AMP kinase. [europepmc]
• Regulation of NAD(P)H oxidases by AMPK in cardiovascular systems. [europepmc]
• Regulation of eNOS enzyme activity by posttranslational modification. [europepmc]
• AMPK: a cellular metabolic and redox sensor. A minireview. [europepmc]
• Shear-induced endothelial mechanotransduction: the interplay between reactive oxygen species (ROS) and nitric oxide (NO) and the pathophysiological implications. [europepmc]
• Cellular and molecular biology of aging endothelial cells. [europepmc]
• Differential Role of Leptin and Adiponectin in Cardiovascular System. [europepmc]
• Endothelial nitric oxide synthase: From biochemistry and gene structure to clinical implications of NOS3 polymorphisms. [europepmc]
• Endothelial nitric oxide synthase: a potential therapeutic target for cerebrovascular diseases. [europepmc]
• AMP-Activated Protein Kinase: An Ubiquitous Signaling Pathway With Key Roles in the Cardiovascular System. [europepmc]
• Mechanisms of Dysfunction in the Aging Vasculature and Role in Age-Related Disease. [europepmc]
• Mechanisms of Vascular Aging. [europepmc]
• New insights into oxidative stress and inflammation during diabetes mellitus-accelerated atherosclerosis. [europepmc]
• PRKAA1/AMPKα1-driven glycolysis in endothelial cells exposed to disturbed flow protects against atherosclerosis. [europepmc]
• AMPK: a therapeutic target of heart failure-not only metabolism regulation. [europepmc]
• AMPK, Mitochondrial Function, and Cardiovascular Disease. [europepmc]
• SIRT1 Antagonizes Oxidative Stress in Diabetic Vascular Complication. [europepmc]

Related

• Shear stress-stimulated AMPK couples endothelial cell mechanics, metabolism and vasodilation
• Thirty Years of Saying NO
• Protein phosphatase 4 mediates palmitic acid-induced endothelial dysfunction by decreasing eNOS phosphorylation at serine 633 in HUVECs
• Cellular and molecular mechanisms of endothelial cell dysfunction.
• eNOS uncoupling and endothelial dysfunction in aged vessels