Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells. Mol Biol Cell 2018 May 15;29(10):1190-1202
Date
03/23/2018Pubmed ID
29563255Pubmed Central ID
PMC5935069DOI
10.1091/mbc.E17-01-0049Scopus ID
2-s2.0-85047085775 (requires institutional sign-in at Scopus site) 97 CitationsAbstract
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 Ca2+ 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.
Author List
Chen Z, D S Oliveira S, Zimnicka AM, Jiang Y, Sharma T, Chen S, Lazarov O, Bonini MG, Haus JM, Minshall RDMESH terms used to index this publication - Major topics in bold
AdultAlbumins
Biopsy
Calcimycin
Calcium
Case-Control Studies
Caveolin 1
Cell Membrane
Diabetes Mellitus, Type 2
Endothelial Cells
HEK293 Cells
Human Umbilical Vein Endothelial Cells
Humans
Insulin
Intercellular Junctions
Ionophores
Middle Aged
Molecular Weight
Nitric Oxide
Nitric Oxide Synthase Type III
Nitrosation
Phosphorylation
Protein Transport
RNA, Small Interfering
src-Family Kinases
