Involvement of reactive oxygen species in a feed-forward mechanism of Na/K-ATPase-mediated signaling transduction

J Biol Chem. 2013 Nov 22;288(47):34249-34258. doi: 10.1074/jbc.M113.461020. Epub 2013 Oct 11.


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.

Keywords: Hypertension; NAK-ATPase; Na/K-ATPase Signaling; Reactive Oxygen Species (ROS); Signal Transduction; Sodium Transport.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetylcysteine / pharmacology
  • Animals
  • CSK Tyrosine-Protein Kinase
  • Cell Line
  • Enzyme Inhibitors / pharmacology
  • Free Radical Scavengers / pharmacokinetics
  • Multienzyme Complexes / genetics
  • Multienzyme Complexes / metabolism*
  • Ouabain / pharmacology
  • Protein Carbonylation / drug effects
  • Protein Carbonylation / genetics
  • Proteolysis / drug effects
  • Reactive Oxygen Species / metabolism*
  • Signal Transduction / drug effects
  • Signal Transduction / physiology*
  • Sodium-Potassium-Exchanging ATPase / genetics
  • Sodium-Potassium-Exchanging ATPase / metabolism*
  • Swine
  • src-Family Kinases


  • Enzyme Inhibitors
  • Free Radical Scavengers
  • Multienzyme Complexes
  • Reactive Oxygen Species
  • Ouabain
  • CSK Tyrosine-Protein Kinase
  • src-Family Kinases
  • Sodium-Potassium-Exchanging ATPase
  • Acetylcysteine