| 19542013 |
Figtree GA, Liu CC, Bibert S, Hamilton EJ, Garcia A, White CN, Chia KK, Cornelius F, Geering K, Rasmussen HH: Reversible oxidative modification: a key mechanism of Na+-K+ pump regulation. Circ Res. 2009 Jul 17;105(2):185-93. Epub 2009 Jun 18.
Angiotensin II (Ang II) inhibits the cardiac sarcolemmal Na (+)-K (+) pump via protein kinase (PK) C-dependent activation of NADPH oxidase. We examined whether this is mediated by oxidative modification of the pump subunits. We detected glutathionylation of beta (1), but not alpha (1), subunits in rabbit ventricular myocytes at baseline. beta (1) Subunit glutathionylation was increased by peroxynitrite (ONOO (-)), paraquat, or activation of NADPH oxidase by Ang II. Increased glutathionylation was associated with decreased alpha (1)/beta (1) subunit coimmunoprecipitation. Glutathionylation was reversed after addition of superoxide dismutase. Glutaredoxin 1, which catalyzes deglutathionylation, coimmunoprecipitated with beta (1) subunit and, when included in patch pipette solutions, abolished paraquat-induced inhibition of myocyte Na (+)-K (+) pump current (I (p)). Cysteine (Cys46) of the beta (1) subunit was the likely candidate for glutathionylation. We expressed Na (+)-K (+) pump alpha (1) subunits with wild-type or Cys46-mutated beta (1) subunits in Xenopus oocytes. ONOO (-) induced glutathionylation of beta (1) subunit and a decrease in Na (+)-K (+) pump turnover number. This was eliminated by mutation of Cys46. ONOO (-) also induced glutathionylation of the Na (+)-K (+) ATPase beta (1) subunit from pig kidney. This was associated with a approximately 2-fold decrease in the rate-limiting E (2)--> E (1) conformational change of the pump, as determined by RH421 fluorescence. We propose that kinase-dependent regulation of the Na (+)-K (+) pump occurs via glutathionylation of its beta (1) subunit at Cys46. These findings have implications for pathophysiological conditions characterized by neurohormonal dysregulation, myocardial oxidative stress and raised myocyte Na (+) levels. |
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