Peroxiredoxin 6 Phosphorylation and Subsequent Phospholipase A2 Activity Are Required for Agonist-mediated Activation of NADPH Oxidase in Mouse Pulmonary Microvascular Endothelium and Alveolar Macrophages
| Autor: | Shampa Chatterjee, David W. Speicher, Kris DeBolt, Aron B. Fisher, Su Nguyen, Elena M. Sorokina, Chandra Dodia, Yu-Chin Lien, Sheldon I. Feinstein |
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| Rok vydání: | 2011 |
| Předmět: |
rac1 GTP-Binding Protein
Mutation Missense Biochemistry Mice Enzyme activator Cytosol Phospholipase A2 Macrophages Alveolar Nitriles Butadienes Null cell Animals Endothelium Enzyme Inhibitors Phosphorylation Lung Molecular Biology Mitogen-Activated Protein Kinase Kinases Oxidase test Membrane Glycoproteins NADPH oxidase biology Angiotensin II Cell Membrane Neuropeptides NADPH Oxidases Cell Biology Molecular biology Mice Mutant Strains rac GTP-Binding Proteins Enzyme Activation Endothelial stem cell Phospholipases A2 Protein Transport Amino Acid Substitution NADPH Oxidase 2 Carcinogens biology.protein Tetradecanoylphorbol Acetate lipids (amino acids peptides and proteins) Reactive Oxygen Species Signal Transduction Peroxiredoxin VI |
| Zdroj: | Journal of Biological Chemistry. 286:11696-11706 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m110.206623 |
| Popis: | Peroxiredoxin 6 (Prdx6), a bifunctional enzyme with glutathione peroxidase and phospholipase A2 (PLA(2)) activities, participates in the activation of NADPH oxidase 2 (NOX2) in neutrophils, but the mechanism for this effect is not known. We now demonstrate that Prdx6 is required for agonist-induced NOX2 activation in pulmonary microvascular endothelial cells (PMVEC) and that the effect requires the PLA(2) activity of Prdx6. Generation of reactive oxygen species (ROS) in response to angiotensin II (Ang II) or phorbol 12-myristate 13-acetate was markedly reduced in perfused lungs and isolated PMVEC from Prdx6 null mice. Rac1 and p47(phox), cytosolic components of NOX2, translocated to the endothelial cell membrane after Ang II treatment in wild-type but not Prdx6 null PMVEC. MJ33, an inhibitor of Prdx6 PLA(2) activity, blocked agonist-induced PLA(2) activity and ROS generation in PMVEC by80%, whereas inhibitors of other PLA(2)s were ineffective. Transfection of Prx6 null cells with wild-type and C47S mutant Prdx6, but not with mutants of the PLA(2) active site (S32A, H26A, and D140A), "rescued" Ang II-induced PLA(2) activity and ROS generation. Ang II treatment of wild-type cells resulted in phosphorylation of Prdx6 and its subsequent translocation from the cytosol to the cell membrane. Phosphorylation as well as PLA(2) activity and ROS generation were markedly reduced by the MAPK inhibitor, U0126. Thus, agonist-induced MAPK activation leads to Prdx6 phosphorylation and translocation to the cell membrane, where its PLA(2) activity facilitates assembly of the NOX2 complex and activation of the oxidase. |
| Databáze: | OpenAIRE |
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