Regulation of endothelial nitric oxide synthase by the actin cytoskeleton

Yunchao Su, Sophia Edwards-Bennett, Michael R. Bubb, Edward R. Block

Research output: Contribution to journalArticle

80 Citations (Scopus)

Abstract

In the present study, the association of endothelial nitric oxide synthase (eNOS) with the actin cytoskeleton in pulmonary artery endothelial cells (PAEC) was examined. We found that the protein contents of eNOS, actin, and caveolin-1 were significantly higher in the caveolar fraction of plasma membranes than in the noncaveolar fraction of plasma membranes in PAEC. Immunoprecipitation of eNOS from lysates of caveolar fractions of plasma membranes in PAEC resulted in the coprecipitation of actin, and immunoprecipitation of actin from lysates of caveolar fractions resulted in the coprecipitation of eNOS. Confocal microscopy of PAEC, in which eNOS was labeled with fluorescein, F-actin was labeled with Texas red-phalloidin, and G-actin was labeled with deoxyribonuclease I conjugated with Texas red, also demonstrated an association between eNOS and F-actin or G-actin. Incubation of purified eNOS with purified F-actin and G-actin resulted in an increase in eNOS activity. The increase in eNOS activity caused by G-actin was much higher than that caused by F-actin. Incubation of PAEC with swinholide A, an actin filament disruptor, resulted in an increase in eNOS activity, eNOS protein content, and association of eNOS with G-actin and in a decrease in the association of eNOS with F-actin. The increase in eNOS activity was higher than that in eNOS protein content in swinholide A-treated cells. In contrast, exposure of PAEC to phalloidin, an actin filament stabilizer, caused decreases in eNOS activity and association of eNOS with G-actin and increases in association of eNOS with F-actin. These results suggest that eNOS is associated with actin in PAEC and that actin and its polymerization state play an important role in the regulation of eNOS activity.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Cell Physiology
Volume284
Issue number6 53-6
StatePublished - Jun 1 2003
Externally publishedYes

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Nitric Oxide Synthase Type III
Actin Cytoskeleton
Actins
Endothelial cells
Pulmonary Artery
Endothelial Cells
Association reactions
Cell membranes
Phalloidine
Cell Membrane
Coprecipitation
Immunoprecipitation
Caveolin 1
Proteins
Confocal microscopy

Keywords

  • Caveolae
  • Endothelial nitric oxide synthase
  • Endothelium
  • Nitric oxide

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology

Cite this

Regulation of endothelial nitric oxide synthase by the actin cytoskeleton. / Su, Yunchao; Edwards-Bennett, Sophia; Bubb, Michael R.; Block, Edward R.

In: American Journal of Physiology - Cell Physiology, Vol. 284, No. 6 53-6, 01.06.2003.

Research output: Contribution to journalArticle

Su, Yunchao ; Edwards-Bennett, Sophia ; Bubb, Michael R. ; Block, Edward R. / Regulation of endothelial nitric oxide synthase by the actin cytoskeleton. In: American Journal of Physiology - Cell Physiology. 2003 ; Vol. 284, No. 6 53-6.
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