Modulation of sarcoplasmic reticulum Ca2+-ATPase by chronic and acute exposure to peroxynitrite

Yolanda Gutiérrez-Martín, Francisco J. Martín-Romero, Francisco A. Iñesta-Vaquera, Carlos Gutiérrez-Merino, Fernando Henao

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Abstract

The Ca2+-ATPase of skeletal muscle sarcoplasmic reticulum (SERCA), an integral membrane protein, becomes irreversibly inactivated in vitro by the addition of a single bolus of peroxynitrite with a K0.5 of 200-300 μM, and this results in a large decrease of the ATP-dependent Ca 2+ gradient across the sarcoplasmic reticulum (SR) membranes. The inactivation of SERCA is raised by treatment of SR vesicles with repetitive micromolar pulses of peroxynitrite. The inhibition of the SERCA is due to the oxidation of thiol groups and tyrosine nitration. Scavengers that react directly with peroxynitrite, such as cysteine, reduced glutathione, NADH, methionine, ascorbate or Trolox, a water-soluble analog of α-tocopherol, afforded significant protection. However, dimethyl sulfoxide and mannitol, two hydroxyl radical scavengers, and α-tocopherol did not protect SERCA from inactivation. Our results showed that the target of peroxynitrite is the cytosolic globular domain of the SERCA and that major skeletal muscle intracellular reductants (ascorbate, NADH and reduced glutathione) protected against inhibition of this ATPase by peroxynitrite.

Original languageEnglish
Pages (from-to)2647-2657
Number of pages11
JournalEuropean Journal of Biochemistry
Volume271
Issue number13
DOIs
Publication statusPublished - Jul 2004

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Keywords

  • Ca-ATPase
  • Calcium homeostasis
  • Peroxynitrite
  • Sarcoplasmic reticulum

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