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Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells
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...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The American Society for Cell Biology
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5935069/ https://www.ncbi.nlm.nih.gov/pubmed/29563255 http://dx.doi.org/10.1091/mbc.E17-01-0049 |
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author | Chen, Zhenlong D. S. Oliveira, Suellen Zimnicka, Adriana M. Jiang, Ying Sharma, Tiffany Chen, Stone Lazarov, Orly Bonini, Marcelo G. Haus, Jacob M. Minshall, Richard D. |
author_facet | Chen, Zhenlong D. S. Oliveira, Suellen Zimnicka, Adriana M. Jiang, Ying Sharma, Tiffany Chen, Stone Lazarov, Orly Bonini, Marcelo G. Haus, Jacob M. Minshall, Richard D. |
author_sort | Chen, Zhenlong |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5935069 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | The American Society for Cell Biology |
record_format | MEDLINE/PubMed |
spelling | pubmed-59350692018-07-30 Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells Chen, Zhenlong D. S. Oliveira, Suellen Zimnicka, Adriana M. Jiang, Ying Sharma, Tiffany Chen, Stone Lazarov, Orly Bonini, Marcelo G. Haus, Jacob M. Minshall, Richard D. Mol Biol Cell Articles 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. The American Society for Cell Biology 2018-05-15 /pmc/articles/PMC5935069/ /pubmed/29563255 http://dx.doi.org/10.1091/mbc.E17-01-0049 Text en © 2018 Chen et al. “ASCB®,” “The American Society for Cell Biology®,” and “Molecular Biology of the Cell®” are registered trademarks of The American Society for Cell Biology. http://creativecommons.org/licenses/by-nc-sa/3.0/ This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License. |
spellingShingle | Articles Chen, Zhenlong D. S. Oliveira, Suellen Zimnicka, Adriana M. Jiang, Ying Sharma, Tiffany Chen, Stone Lazarov, Orly Bonini, Marcelo G. Haus, Jacob M. Minshall, Richard D. Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells |
title | Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells |
title_full | Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells |
title_fullStr | Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells |
title_full_unstemmed | Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells |
title_short | Reciprocal regulation of eNOS and caveolin-1 functions in endothelial cells |
title_sort | reciprocal regulation of enos and caveolin-1 functions in endothelial cells |
topic | Articles |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5935069/ https://www.ncbi.nlm.nih.gov/pubmed/29563255 http://dx.doi.org/10.1091/mbc.E17-01-0049 |
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