Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells

Zhenlong Chen, Suellen D.S. Oliveira, Adriana M. Zimnicka, Ying Jiang, Tiffany Sharma, Stone Chen, Orly Lazarov, Marcelo G. Bonini, Jacob M. Haus, Richard D. Minshall*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

58 Scopus citations


We hypothesized that the maintenance of vascular homeostasis is critically dependent on the expression and reciprocal regulation of caveolin-1 (Cav-1) and endothelial nitric oxide synthase (eNOS) in endothelial cells (ECs). Skeletal muscle biopsies from subjects with type 2 diabetes showed 50% less Cav-1 and eNOS than those from lean healthy controls. The Cav-1:eNOS expression ratio was 200:1 in primary culture human ECs. Cav-1 small interfering RNA (siRNA) reduced eNOS protein and gene expression in association with a twofold increase in eNOS phosphorylation and nitrate production per molecule of eNOS, which was reversed in cells overexpressing Adv-Cav-1-GFP. Upon addition of the Ca 2+ ionophore A23187 to activate eNOS, we observed eNOS Ser1177 phosphorylation, its translocation to β-catenin-positive cell–cell junctions, and increased colocalization of eNOS and Cav-1 within 5 min. We also observed Cav-1 S-nitrosylation and destabilization of Cav-1 oligomers in cells treated with A23187 as well as insulin or albumin, and this could be blocked by L-NAME, PP2, or eNOS siRNA. Finally, caveola-mediated endocytosis of albumin or insulin was reduced by Cav-1 or eNOS siRNA, and the effect of Cav-1 siRNA was rescued by Adv-Cav-1-GFP. Thus, Cav-1 stabilizes eNOS expression and regulates its activity, whereas eNOS-derived NO promotes caveola-mediated endocytosis.

Original languageEnglish (US)
Pages (from-to)1190-1202
Number of pages13
JournalMolecular biology of the cell
Issue number10
StatePublished - May 15 2018
Externally publishedYes

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology


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