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Nitric Oxide‐mediated Proteins' S‐nitrosation in Endothelial Cells under Shear Flow or Rosuvastatin Treatment
Author(s) -
Wang Ling D.,
Huang Bin
Publication year - 2010
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.24.1_supplement.984.13
Subject(s) - chemistry , nitric oxide , nitrosation , enos , protein disulfide isomerase , western blot , hsp70 , heat shock protein , rosuvastatin , statin , biochemistry , microbiology and biotechnology , pharmacology , biology , nitric oxide synthase , enzyme , organic chemistry , gene
Atheroprotection of shear flow to endothelial cells (ECs) is partly contributed by the increased nitric oxide (NO) production. Cholesterol lowing drug, statin, is known to increase NO levels via the induction of eNOS expression and activity. NO‐mediated S‐nitrosation of Cys residue has emerged as a fundamental post‐translational modification of proteins in ECs. S‐nitrosation can alter protein's function and affects vascular physiology. In the present study, proteins' S‐nitrosation was examined in HUVECs after shear flow or statin treament. Increased eNOS activation was observed in ECs after rosuvastatin (10 μM) or shear flow (25 dynes/cm2) treatment. The enhanced S‐nitrosoproteins in rosuvastatin‐treated ECs were detected by biotin switch method coupled with Western blot‐based 2‐DE, while the shear flow‐enhanced S‐nitrosated proteins were detected by CyDye DIGE. There were 12 and 18 proteins showing increased S‐nitrosation in ECs under flow and statin treatments, respectively. S‐nitrosation in protein disulfide isomerase (PDI), mitochondrial heat shock protein 70 (mt‐HSP70), heat shock transcription factor, tropomyosin and vimentin were shown to be increased in both treatments. Further analysis by nLC‐MS/MS and MASCOT in‐house algorithm, the S‐nitrosated Cys residues on the mt‐HSP70, tropomyosin and vimentin were also determined. Our results indicating that statin or shear flow exerts similar atheroprotection to ECs is at least partly due to the NO‐mediated S‐nitrosation in proteins. Proteins' S‐nitrosation may be essential for maintaining endothelial integrity. (This work ws supported by National Science Council and Academia Sinica, Taiwan)