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Regulation of the ADMA/PRMT/DDAH pathway in vascular smooth muscle cells by the p22phox component of NADPH oxidase
Author(s) -
Luo Zaiming,
Welch William,
Teerlink Tom,
Griendling Kathy,
Wilcox Christopher
Publication year - 2009
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.23.1_supplement.803.2
Subject(s) - asymmetric dimethylarginine , arginine , nadph oxidase , p22phox , medicine , reactive oxygen species , western blot , protein arginine methyltransferase 5 , vascular smooth muscle , chemistry , endocrinology , biochemistry , biology , methyltransferase , methylation , amino acid , gene , smooth muscle
We reported increased reactive oxygen species (ROS) and asymmetric dimethylarginine (ADMA) in chronic kidney disease or hypertension. Arginine is methylated by protein arginine methyltransferases (PRMTs). ADMA is metabolized by dimethylarginine dimethylaminohydrolases (DDAHs). We tested the hypothesis that ROS dictate cellular arginine methylation. DDAH‐1 and ‐2 protein expression were increased by 37 ± 6 and 50 ± 4% (p<0.05) and PRMT‐3 protein increased 41 ± 7%(p< 0.05) in aortas of mice transgenic for p22 phox in VSMCs (Tg p22smc ), despite unchanged plasma ADMA (0.59 ± 0.1 vs 0.56 ± 0.1 μmol‐ L −1 ; ns). Rat preglomerular vascular smooth muscle cells (PGVSMCs) stably transfected with p22 phox (S‐p22 phox ) which generated two‐fold more ROS had a 62± 2% (p<0.01) reduced DDAH activity, and a 51 ± 5% and 58 ± 4% (both p<0.05) reduced protein, but increased mRNA expression, for DDAH‐1 and ‐2. Western blots demonstrated substantial, but selective, protein carbonylation and asymmetric dimethylation of arginine moieties in lysates from S‐p22 phox cells that were prevented by PEG‐catalase (500 units · ml −1 ) or tempol (10 −4 M). In conclusion, ROS produced by NADPH oxidase in VSMCs enhances PRMT and cellular accumulation of arginine methylated proteins. Protein carbonylation may contribute to decreased DDAH protein expression, despite increased mRNA. The similar changes in blood vessel of Tg p22smc mice demonstrates that these effects could underlie endothelial dysfunction.