Coordinated regulation of endothelial nitric oxide synthase activity by phosphorylation and subcellular localization

Yong Chool Boo, Hyo Jung Kim, Hannah Song, David J Fulton, William Sessa, Hanjoong Jo

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Endothelial nitric oxide synthase (eNOS) is regulated by multiple mechanisms including Ca2+/calmodulin binding, protein-protein interactions, phosphorylation, and subcellular locations. Emerging evidence suggests that these seemingly independent mechanisms may be closely correlated. In the present study, the interplay between membrane targeting and phosphorylation of eNOS was investigated by using various mutants designed to target specific subcellular locations or to mimic different phospho states. Phospho-mimicking mutations of wild-type eNOS at S635 and S1179 synergistically activated the enzyme. The targeted eNOS mutants to plasma membrane and Golgi complex exhibited higher NO production activities than that of a myristoylation-deficient cytosolic mutant. Phospho-mimicking mutations at S635 and S1179 rescued the activity of the cytosolic mutant and increased those of the plasma membrane- and Golgi-targeted mutants. In contrast, phospho-deficient mutations at these sites led to inactivation of eNOS. Unlike the other targeted mutants, the cytosolic eNOS mutant was unresponsive to cAMP, indicating that membrane association and phosphorylation are required for eNOS activation. These findings suggest that the coordinated interplay between phosphorylation and subcellular localization of eNOS plays an important role in regulating NO production in endothelial cells.

Original languageEnglish (US)
Pages (from-to)144-153
Number of pages10
JournalFree Radical Biology and Medicine
Volume41
Issue number1
DOIs
StatePublished - Jul 1 2006

Fingerprint

Phosphorylation
Nitric Oxide Synthase Type III
Cell membranes
Mutation
Cell Membrane
Calmodulin-Binding Proteins
Membranes
Endothelial cells
Golgi Apparatus
Endothelial Cells
Chemical activation
Association reactions
Enzymes

Keywords

  • Free radical
  • Membrane localization
  • Phosphorylation
  • eNOS mutants

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

Cite this

Coordinated regulation of endothelial nitric oxide synthase activity by phosphorylation and subcellular localization. / Boo, Yong Chool; Kim, Hyo Jung; Song, Hannah; Fulton, David J; Sessa, William; Jo, Hanjoong.

In: Free Radical Biology and Medicine, Vol. 41, No. 1, 01.07.2006, p. 144-153.

Research output: Contribution to journalArticle

Boo, Yong Chool ; Kim, Hyo Jung ; Song, Hannah ; Fulton, David J ; Sessa, William ; Jo, Hanjoong. / Coordinated regulation of endothelial nitric oxide synthase activity by phosphorylation and subcellular localization. In: Free Radical Biology and Medicine. 2006 ; Vol. 41, No. 1. pp. 144-153.
@article{d87bf89e84b140dda1be67a6db898e49,
title = "Coordinated regulation of endothelial nitric oxide synthase activity by phosphorylation and subcellular localization",
abstract = "Endothelial nitric oxide synthase (eNOS) is regulated by multiple mechanisms including Ca2+/calmodulin binding, protein-protein interactions, phosphorylation, and subcellular locations. Emerging evidence suggests that these seemingly independent mechanisms may be closely correlated. In the present study, the interplay between membrane targeting and phosphorylation of eNOS was investigated by using various mutants designed to target specific subcellular locations or to mimic different phospho states. Phospho-mimicking mutations of wild-type eNOS at S635 and S1179 synergistically activated the enzyme. The targeted eNOS mutants to plasma membrane and Golgi complex exhibited higher NO production activities than that of a myristoylation-deficient cytosolic mutant. Phospho-mimicking mutations at S635 and S1179 rescued the activity of the cytosolic mutant and increased those of the plasma membrane- and Golgi-targeted mutants. In contrast, phospho-deficient mutations at these sites led to inactivation of eNOS. Unlike the other targeted mutants, the cytosolic eNOS mutant was unresponsive to cAMP, indicating that membrane association and phosphorylation are required for eNOS activation. These findings suggest that the coordinated interplay between phosphorylation and subcellular localization of eNOS plays an important role in regulating NO production in endothelial cells.",
keywords = "Free radical, Membrane localization, Phosphorylation, eNOS mutants",
author = "Boo, {Yong Chool} and Kim, {Hyo Jung} and Hannah Song and Fulton, {David J} and William Sessa and Hanjoong Jo",
year = "2006",
month = "7",
day = "1",
doi = "10.1016/j.freeradbiomed.2006.03.024",
language = "English (US)",
volume = "41",
pages = "144--153",
journal = "Free Radical Biology and Medicine",
issn = "0891-5849",
publisher = "Elsevier Inc.",
number = "1",

}

TY - JOUR

T1 - Coordinated regulation of endothelial nitric oxide synthase activity by phosphorylation and subcellular localization

AU - Boo, Yong Chool

AU - Kim, Hyo Jung

AU - Song, Hannah

AU - Fulton, David J

AU - Sessa, William

AU - Jo, Hanjoong

PY - 2006/7/1

Y1 - 2006/7/1

N2 - Endothelial nitric oxide synthase (eNOS) is regulated by multiple mechanisms including Ca2+/calmodulin binding, protein-protein interactions, phosphorylation, and subcellular locations. Emerging evidence suggests that these seemingly independent mechanisms may be closely correlated. In the present study, the interplay between membrane targeting and phosphorylation of eNOS was investigated by using various mutants designed to target specific subcellular locations or to mimic different phospho states. Phospho-mimicking mutations of wild-type eNOS at S635 and S1179 synergistically activated the enzyme. The targeted eNOS mutants to plasma membrane and Golgi complex exhibited higher NO production activities than that of a myristoylation-deficient cytosolic mutant. Phospho-mimicking mutations at S635 and S1179 rescued the activity of the cytosolic mutant and increased those of the plasma membrane- and Golgi-targeted mutants. In contrast, phospho-deficient mutations at these sites led to inactivation of eNOS. Unlike the other targeted mutants, the cytosolic eNOS mutant was unresponsive to cAMP, indicating that membrane association and phosphorylation are required for eNOS activation. These findings suggest that the coordinated interplay between phosphorylation and subcellular localization of eNOS plays an important role in regulating NO production in endothelial cells.

AB - Endothelial nitric oxide synthase (eNOS) is regulated by multiple mechanisms including Ca2+/calmodulin binding, protein-protein interactions, phosphorylation, and subcellular locations. Emerging evidence suggests that these seemingly independent mechanisms may be closely correlated. In the present study, the interplay between membrane targeting and phosphorylation of eNOS was investigated by using various mutants designed to target specific subcellular locations or to mimic different phospho states. Phospho-mimicking mutations of wild-type eNOS at S635 and S1179 synergistically activated the enzyme. The targeted eNOS mutants to plasma membrane and Golgi complex exhibited higher NO production activities than that of a myristoylation-deficient cytosolic mutant. Phospho-mimicking mutations at S635 and S1179 rescued the activity of the cytosolic mutant and increased those of the plasma membrane- and Golgi-targeted mutants. In contrast, phospho-deficient mutations at these sites led to inactivation of eNOS. Unlike the other targeted mutants, the cytosolic eNOS mutant was unresponsive to cAMP, indicating that membrane association and phosphorylation are required for eNOS activation. These findings suggest that the coordinated interplay between phosphorylation and subcellular localization of eNOS plays an important role in regulating NO production in endothelial cells.

KW - Free radical

KW - Membrane localization

KW - Phosphorylation

KW - eNOS mutants

UR - http://www.scopus.com/inward/record.url?scp=33745080255&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33745080255&partnerID=8YFLogxK

U2 - 10.1016/j.freeradbiomed.2006.03.024

DO - 10.1016/j.freeradbiomed.2006.03.024

M3 - Article

VL - 41

SP - 144

EP - 153

JO - Free Radical Biology and Medicine

JF - Free Radical Biology and Medicine

SN - 0891-5849

IS - 1

ER -