Caveolin 1 is required for the activation of endothelial nitric oxide synthase in response to 17β-estradiol

Neetu Sud, Dean A. Wiseman, Stephen Matthew Black

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Evidence suggests that estrogen mediates rapid endothelial nitric oxide synthase (eNOS) activation via estrogen receptor-a (ERα) within the plasma membrane of endothelial cells (EC). ERα is known to colocalize with caveolin 1, the major structural protein of caveolae, and caveolin 1 stimulates the translocation of ERα to the plasma membrane. However, the role played by caveolin 1 in regulating 17β-estradiol-mediated NO signaling in EC has not been adequately resolved. Thus, the purpose of this study was to explore how 17β-estradiol stimulates eNOS activity and the role of caveolin 1 in this process. Our data demonstrate that modulation of caveolin 1 expression using small interfering RNA or adenoviral gene delivery alters ERα localization to the plasma membrane in EC. Further, before estrogen stimulation ERα associates with caveolin 1, whereas stimulation promotes a pp60 Src-mediated phosphorylation of caveolin 1 at tyrosine 14, increasing ERα-PI3 kinase interactions and disrupting caveolin 1-ERα interactions. Adenoviral mediated over-expression of a phosphorylation-deficient mutant of caveolin (Y14FCav) attenuated the ERα/PI3 kinase interaction and prevented Akt-mediated eNOS activation. Furthermore, Y14FCav over-expression reduced eNOS phosphorylation at serine 1177 and decreased NO generation after estrogen exposure. Using a library of overlapping peptides we identified residues 62-73 of caveolin 1 as the ERα-binding site. Delivery of a synthetic peptide based on this sequence decreased ERα plasma membrane translocation and reduced estrogen-mediated activation of eNOS. In conclusion, caveolin 1 stimulates 17β-estradiol-induced NO production by promoting ERα to the plasma membrane, which facilitates the activation of the PI3 kinase pathway, leading to eNOS activation and NO generation.

Original languageEnglish (US)
Pages (from-to)1637-1649
Number of pages13
JournalMolecular Endocrinology
Volume24
Issue number8
DOIs
StatePublished - Aug 1 2010

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Caveolin 1
Nitric Oxide Synthase Type III
Estrogen Receptors
Estradiol
Cell Membrane
Estrogens
Phosphatidylinositol 3-Kinases
Endothelial Cells
Phosphorylation
Caveolins
Peptide Library
Caveolae
Serine
Small Interfering RNA
Tyrosine
Binding Sites

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology

Cite this

Caveolin 1 is required for the activation of endothelial nitric oxide synthase in response to 17β-estradiol. / Sud, Neetu; Wiseman, Dean A.; Black, Stephen Matthew.

In: Molecular Endocrinology, Vol. 24, No. 8, 01.08.2010, p. 1637-1649.

Research output: Contribution to journalArticle

Sud, Neetu ; Wiseman, Dean A. ; Black, Stephen Matthew. / Caveolin 1 is required for the activation of endothelial nitric oxide synthase in response to 17β-estradiol. In: Molecular Endocrinology. 2010 ; Vol. 24, No. 8. pp. 1637-1649.
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