title: Involvement of reactive oxygen species in a feed-forward mechanism of Na/K-ATPase-mediated signaling transduction.
abstract: Cardiotonic steroids (such as ouabain) signaling through Na/K-ATPase regulate sodium reabsorption in the renal proximal tubule. We report here that reactive oxygen species are required to initiate ouabain-stimulated Na/K-ATPase·c-Src signaling. Pretreatment with the antioxidant N-acetyl-L-cysteine prevented ouabain-stimulated Na/K-ATPase·c-Src signaling, protein carbonylation, redistribution of Na/K-ATPase and sodium/proton exchanger isoform 3, and inhibition of active transepithelial (22)Na(+) transport. Disruption of the Na/K-ATPase·c-Src signaling complex attenuated ouabain-stimulated protein carbonylation. Ouabain-stimulated protein carbonylation is reversed after removal of ouabain, and this reversibility is largely independent of de novo protein synthesis and degradation by either the lysosome or the proteasome pathways. Furthermore, ouabain stimulated direct carbonylation of two amino acid residues in the actuator domain of the Na/K-ATPase α1 subunit. Taken together, the data indicate that carbonylation modification of the Na/K-ATPase α1 subunit is involved in a feed-forward mechanism of regulation of ouabain-mediated renal proximal tubule Na/K-ATPase signal transduction and subsequent sodium transport.
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, JCE School of Medicine at Marshall University, Huntington, West Virginia 25755; Institute of Biomedical Engineering, Yanshan University, Qinhuangdao 066004, China.
authors.forename: Yanling
authors.initials: Y
authors.lastname: Yan
authors.affiliation: Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Anna P
authors.initials: AP
authors.lastname: Shapiro
authors.affiliation: Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Steven
authors.initials: S
authors.lastname: Haller
authors.affiliation: Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Vinai
authors.initials: V
authors.lastname: Katragadda
authors.affiliation: Department of Pharmacology, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Lijun
authors.initials: L
authors.lastname: Liu
authors.affiliation: Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614; Department of Pharmacology, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Jiang
authors.initials: J
authors.lastname: Tian
authors.affiliation: Department of Pathology, University of Michigan, Ann Arbor, Michigan 48109.
authors.forename: Venkatesha
authors.initials: V
authors.lastname: Basrur
authors.affiliation: Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Deepak
authors.initials: D
authors.lastname: Malhotra
authors.affiliation: Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614; Department of Pharmacology, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Zi-Jian
authors.initials: ZJ
authors.lastname: Xie
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, JCE School of Medicine at Marshall University, Huntington, West Virginia 25755.
authors.forename: Nader G
authors.initials: NG
authors.lastname: Abraham
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, JCE School of Medicine at Marshall University, Huntington, West Virginia 25755; Department of Medicine, University of Toledo College of Medicine, Toledo, Ohio 43614.
authors.forename: Joseph I
authors.initials: JI
authors.lastname: Shapiro
authors.affiliation: Department of Pharmacology, Physiology and Toxicology, JCE School of Medicine at Marshall University, Huntington, West Virginia 25755. Electronic address: liuj@marshall.edu.
authors.forename: Jiang
authors.initials: J
authors.lastname: Liu
mesh_terms: D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D000081247:CSK Tyrosine-Protein Kinase; D002460:Cell Line; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000493:pharmacokinetics; D009097:Multienzyme Complexes / Q000235:genetics / Q000378:metabolism*; D010042:Ouabain / Q000494:pharmacology; D050050:Protein Carbonylation / Q000187:drug effects / Q000235:genetics; D059748:Proteolysis / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology*; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism*; D013552:Swine; D019061:src-Family Kinases
chemical_list: D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D009097:Multienzyme Complexes; D017382:Reactive Oxygen Species; D010042:Ouabain; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; D000254:Sodium-Potassium-Exchanging ATPase; D000111:Acetylcysteine
keywords: Hypertension; NAK-ATPase; Na/K-ATPase Signaling; Reactive Oxygen Species (ROS); Signal Transduction; Sodium Transport