+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Localization of endothelial nitric-oxide synthase phosphorylated on serine 1179 and nitric oxide in Golgi and plasma membrane defines the existence of two pools of active enzyme



Localization of endothelial nitric-oxide synthase phosphorylated on serine 1179 and nitric oxide in Golgi and plasma membrane defines the existence of two pools of active enzyme



Journal of Biological Chemistry 277(6): 4277-4284



The subcellular localization of endothelial nitric-oxide synthase (eNOS) is critical for optimal coupling of extracellular stimulation to nitric oxide production. Because eNOS is activated by Akt-dependent phosphorylation to produce nitric oxide (NO), we determined the subcellular distribution of eNOS phosphorylated on serine 1179 using a variety of methodologies. Based on sucrose gradient fractionation, phosphorylated-eNOS (P-eNOS) was found in both caveolin-1-enriched membranes and intracellular domains. Co-transfection of eNOS with Akt and stimulation of endothelial cells with vascular endothelial growth factor (VEGF) increased the ratio of P-eNOS to total eNOS but did not change the relative intracellular distribution between these domains. The proper localization of eNOS to intracellular membranes was required for agonist-dependent phosphorylation on serine 1179, since VEGF did not increase eNOS phosphorylation in cells transfected with a non-acylated, mistargeted form of eNOS. Confocal imaging of P-eNOS and total eNOS pools demonstrated co-localization in the Golgi region and plasmalemma of transfected cells and native endothelial cells. Finally, VEGF stimulated a large increase in NO localized in both the perinuclear region and the plasma membrane of endothelial cells. Thus, activated, phosphorylated eNOS resides in two cellular compartments and both pools are VEGF-regulated to produce NO.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 010931533

Download citation: RISBibTeXText

PMID: 11729179

DOI: 10.1074/jbc.m106302200


Related references

Localization of Activity-Dependent Neuroprotective Protein-like immunoreactivity in the neonatal rat brain. Society for Neuroscience Abstracts 27(1): 949, 2001

Endothelial cell regulation of matrix metalloproteinases. Canadian Journal of Physiology and Pharmacology 83(1): 1-7, 2005

Knockdown of legumain inhibits cleavage of annexin A2 in the mouse kidney. Biochemical and Biophysical Research Communications 430(2): 482-487, 2013

Complete genome sequencing and evolutionary analysis of Indian isolates of Dengue virus type 2. Biochemical and Biophysical Research Communications 436(3): 478-485, 2013

Targeting of diethylene triamine pentaacetic acid encapsulated in liposomes to rat liver: an effective strategy to prevent bone deposition and increase urine elimination of plutonium in rats. International Journal of Radiation Biology 80(6): 413-422, 2004

Retinoic acid and polyriboinosinic:polyribocytidylic acid stimulate robust anti-tetanus antibody production while differentially regulating type 1/type 2 cytokines and lymphocyte populations. Journal of Immunology 174(12): 7961-7969, 2005

Phosphorylation of retinoid X receptor at serine 260 suppresses its ubiquitin/proteasome-mediated degradation in human hepatocellular carcinoma. Hepatology 34(4 Pt 2): 182A, 2001

Serinc, an activity-regulated protein family, incorporates serine into membrane lipid synthesis. Journal of Biological Chemistry 280(42): 35776-35783, 2005

The profiles of red fluorescent proteins with antinucleopolyhedrovirus activity in races of the silkwormBombyx mori. 2011

Chk1 and Chk2 are differentially involved in homologous recombination repair and cell cycle arrest in response to DNA double-strand breaks induced by camptothecins. Molecular Cancer Therapeutics 7(6): 1440-1449, 2008

Where is child protection in penal custody?. Criminal Behaviour and Mental Health 16(3): 137-141, 2006

Serina Gredler (Gastropoda, Stylommatophora: Enidae), the continuous-peristomed mountain snails endemic to the eastern slope of the Qinghai-Xizang Plateau. Zootaxa 3620: 43-66, 2013

Functional reconstitution of beta-glucan elicitor-binding activity upon incorporation into lipid vesicles. Febs Letters 458(2): 129-132, 1999

Differential Effects of HCN Channel Block on On and Off Pathways in the Retina as a Potential Cause for Medication-Induced Phosphene Perception. Investigative Ophthalmology and Visual Science 58(11): 4754-4767, 2017

Superoxide generation from eosinophils involved translocation of NADPH oxidase components. Journal of Allergy and Clinical Immunology 103(1 Part 2): S111, 1999