S-Glutathiolation by peroxynitrite activates SERCA during arterial relaxation by nitric oxide

Nat Med. 2004 Nov;10(11):1200-7. doi: 10.1038/nm1119. Epub 2004 Oct 17.


Nitric oxide (NO) physiologically stimulates the sarco/endoplasmic reticulum calcium (Ca(2+)) ATPase (SERCA) to decrease intracellular Ca(2+) concentration and relax cardiac, skeletal and vascular smooth muscle. Here, we show that NO-derived peroxynitrite (ONOO(-)) directly increases SERCA activity by S-glutathiolation and that this modification of SERCA is blocked by irreversible oxidation of the relevant cysteine thiols during atherosclerosis. Purified SERCA was S-glutathiolated by ONOO(-) and the increase in Ca(2+)-uptake activity of SERCA reconstituted in phospholipid vesicles required the presence of glutathione. Mutation of the SERCA-reactive Cys674 to serine abolished these effects. Because superoxide scavengers decreased S-glutathiolation of SERCA and arterial relaxation by NO, ONOO(-) is implicated as the intracellular mediator. NO-dependent relaxation as well as S-glutathiolation and activation of SERCA were decreased by atherosclerosis and Cys674 was found to be oxidized to sulfonic acid. Thus, irreversible oxidation of key thiol(s) in disease impairs NO-induced relaxation by preventing reversible S-glutathiolation and activation of SERCA by NO/ONOO(-).

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Arteriosclerosis / metabolism*
  • Calcium / metabolism*
  • Calcium-Transporting ATPases / genetics
  • Calcium-Transporting ATPases / metabolism*
  • Cells, Cultured
  • Chromatography, High Pressure Liquid
  • DNA Primers
  • Glutathione / metabolism*
  • Humans
  • Immunoprecipitation
  • Mass Spectrometry
  • Mice
  • Mice, Transgenic
  • Muscle, Smooth, Vascular / metabolism*
  • Mutagenesis
  • Mutation / genetics
  • Nitric Oxide / metabolism*
  • Peroxynitrous Acid / metabolism*
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases


  • DNA Primers
  • Peroxynitrous Acid
  • Nitric Oxide
  • Sarcoplasmic Reticulum Calcium-Transporting ATPases
  • Calcium-Transporting ATPases
  • Glutathione
  • Calcium