Oxidation of Prx2 and phosphorylation of GRP58 by angiotensin II in human coronary smooth muscle cells identified by 2D-DIGE analysis.
To study early changes in angiotensin II (Ang II)-induced signaling with post-translational modifications, we analyzed proteins from cultured human coronary smooth muscle cells stimulated with Ang II, using two-dimensional difference gel electrophoresis (2D-DIGE) combined with ProQ Diamond and SYPRO Ruby staining, followed by mass spectrometry or Western blotting. Among 40 proteins identified, peroxiredoxin 2 (Prx2) was oxidized and 58 kDa glucose-regulated protein (GRP58) was phosphorylated after 5 min of Ang II (1 microM) stimulation. Valsartan, a selective Ang II type 1 (AT1) receptor blocker, and N-acetylcysteine, an antioxidant, inhibited both of these modifications, indicating the contribution of AT1 receptor and reactive oxygen species to oxidation of Prx2 and phosphorylation of GRP58 by Ang II.
Tokutomi
Yoshiko
Y
Department of Pharmacology and Molecular Therapeutics, Kumamoto University Graduate School of Medical Sciences, Honjo 1-1-1, Kumamoto 860-8556, Japan.
Araki
Norie
N
Kataoka
Keiichiro
K
Yamamoto
Eiichiro
E
Kim-Mitsuyama
Shokei
S
D000804:Angiotensin II / Q000008:administration & dosage*; D002478:Cells, Cultured; D003331:Coronary Vessels; D004305:Dose-Response Relationship, Drug; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D018398:Homeodomain Proteins / Q000378:metabolism*; D006801:Humans; D032389:Myocytes, Smooth Muscle; D010084:Oxidation-Reduction / Q000187:drug effects; D010766:Phosphorylation / Q000187:drug effects; D019704:Protein Disulfide-Isomerases / Q000378:metabolism*
D018398:Homeodomain Proteins; C512693:PRRX2 protein, human; D000804:Angiotensin II; D019704:Protein Disulfide-Isomerases; C085868:PDIA3 protein, human