Shear flow increases S-nitrosylation of proteins in endothelial cells

Bin Huang, Shih Chung Chen, Danny Ling Wang

研究成果: 雜誌貢獻文章同行評審

72 引文 斯高帕斯(Scopus)

摘要

AimsEndothelial cells (ECs) constantly exposed to shear flow increase nitric oxide production via the activation of endothelial nitric oxide synthase. Nitric oxide-mediated S-nitrosylation has recently been identified as an important post-translational modification that may alter signalling and/or protein function. S-nitrosylation of endothelial proteins after shear flow treatment has not been fully explored. In this study, the CyDye switch method was utilized to examine S-nitrosylated proteins in ECs after exposure to shear flow.Methods and resultsHuman umbilical vein ECs were subjected to shear flow for 30 min, and S-nitrosylated proteins were detected by the CyDye switch method. In principle, free thiols in proteins become blocked by alkylation, the S-nitrosylated bond is reduced by ascorbate, and then CyDye labels proteins. Proteins that separately labelled with Cy3 or Cy5 were mixed and subjected to two-dimensional gel electrophoresis for further analysis. More than 100 S-nitrosoproteins were detected in static and shear-treated ECs. Among these, 12 major proteins of heterogeneous function showed a significant increase in S-nitrosylation following shear flow. The S-nitrosylated residues in tropomyosin and vimentin, which were localized in the hydrophobic motif of each protein, were identified as Cys170 and Cys328, respectively.ConclusionPost-translational S-nitrosylation of proteins in ECs can be detected by a reliable CyDye switch method. This flow-induced S-nitrosylation of endothelial proteins may be essential for the adaptation and remodelling of ECs under flow conditions.
原文英語
頁(從 - 到)536-546
頁數11
期刊Cardiovascular Research
83
發行號3
DOIs
出版狀態已發佈 - 8月 2009

ASJC Scopus subject areas

  • 心臟病學與心血管醫學
  • 生理學(醫學)
  • 生理學

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