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Related Experiment Videos

Cytochrome c nitration by peroxynitrite.

A M Cassina1, R Hodara, J M Souza

  • 1Departamento de Bioquimica, Facultad de Medicina and Laboratorio de Enzimologia, Instituto de Quimica Biológica, Facultad de Ciencias, Universidad de la República, 11800 Montevideo, Uruguay.

The Journal of Biological Chemistry
|April 20, 2000
PubMed
Summary
This summary is machine-generated.

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Peroxynitrite nitration damages cytochrome c, a key mitochondrial protein. This damage alters its function, potentially contributing to cell injury during oxidative stress.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Oxidative Stress Research

Background:

  • Peroxynitrite (ONOO(-)), formed from superoxide (O(2)) and nitric oxide (.NO), inhibits mitochondrial respiration and induces apoptosis.
  • Cytochrome c is crucial for mitochondrial respiration and apoptosis, making it a potential target for peroxynitrite during oxidative stress.

Purpose of the Study:

  • To investigate the direct effects of peroxynitrite on horse heart cytochrome c.
  • To determine if peroxynitrite-nitration alters cytochrome c's structure, redox properties, and function.

Main Methods:

  • Incubation of horse heart cytochrome c with peroxynitrite.
  • Spectroscopic analysis (UV-Vis absorption at 695 nm).
  • Western blot analysis using an anti-3-nitrotyrosine antibody.

Related Experiment Videos

  • Mass spectrometry to confirm nitration and identify nitrated residues.
  • Assessment of mitochondrial respiration in rat heart mitochondria.

    • Main Results:

    • Peroxynitrite directly nitrated cytochrome c, primarily at tyrosine 67 (Tyr-67), a heme-vicinal residue.
    • Nitration caused a conformational change, displacing the methionine-80 (Met-80) heme ligand and altering spectral properties.
    • Nitrated cytochrome c exhibited increased peroxidatic activity, resistance to reduction, and impaired support of mitochondrial respiration.

    Conclusions:

    • Cytochrome c is a direct target of peroxynitrite-mediated nitration.
    • Nitration of cytochrome c significantly alters its redox properties and impairs its function in mitochondrial respiration.
    • Peroxynitrite-induced cytochrome c nitration may play a role in oxidative and signaling events during cell injury.