Time Course and Site(s) of Cytochrome c Tyrosine Nitration by Peroxynitrite
Abstract
Cytochrome c-dependent electron transfer and apoptosome activation require protein-protein binding, which are mainly directed by conformational and specific electrostatic interactions. Cytochrome c contains four highly conserved tyrosine residues, one internal (Tyr67), one intermediate (Tyr48), and two more accessible to the solvent (Tyr74 and Tyr97). Tyrosine nitration by biologically-relevant intermediates could influence cytochrome c structure and function. Herein, we analyzed the time course and site(s) of tyrosine nitration in horse cytochrome c by fluxes of peroxynitrite. Also, a method of purifying each (nitrated) cytochrome c product by cation-exchange HPLC was developed. A flux of peroxynitrite caused the time-dependent formation of different nitrated species, all less positively charged than the native form. At low accumulated doses of peroxynitrite, the main products were two mononitrated cytochrome c species at Tyr97 and Tyr74, as shown by peptide mapping and mass spectrometry analysis. At higher doses, all tyrosine residues in cytochrome c were nitrated, including dinitrated (i.e., Tyr97 and Tyr67 or Tyr74 and Tyr67) and trinitrated (i.e., Tyr97, Tyr74, and Tyr67) forms of the protein, with Tyr67 well represented in dinitrated species and Tyr48 being the least prone to nitration. All mono-, di-, and trinitrated cytochrome c species displayed an increased peroxidase activity. Nitrated cytochrome c in Tyr74 and Tyr67, and to a lesser extent in Tyr97, was unable to…
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The heme iron of cytochrome c is predominately hexacoordinate, but can become pentacoordinate when the iron-methionine bond is ruptured which occurs when the methionine is oxidized, tyrosines are nitrated, or the cytochrome interacts with anionic phospholipids.
“Similar to Ngb, the heme iron of cytochrome c is predominately hexacoordinate, but can become pentacoordinate when the iron-methionine bond is ruptured which occurs when the methionine is oxidized [65, 66], tyrosines are nitrated [67, 68], or the cytochrome interacts with anionic phospholipids such as those present in the inner membrane of the mitochondria [69–71].”
From Mechanisms of nitrite bioactivation (2014) · cites this paper as [67]
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References 18 · Cited by 112
- Authors
- Batthyány, C., Souza, J. M., Durán, R., Cassina, A., Cerveñansky, C., & Radi, R.
- Affiliations
- Universidad de la República de Uruguay (UY)
- Departamento de Bioquímica and Center for Free Radical and Biomedical Research, Facultad de Medicina, Universidad de la República, Montevideo, Uruguay, and Unidad de Bioquímica Analítica, Instituto de Investigaciones Biológicas Clemente Estable, Montevideo, Uruguay
- Venue
- Biochemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2005-05-12
- Volume
- 44
- Issue
- 22
- Pages
- 8038-8046
- ISSN
- 0006-2960 (ISSN-L)
- 1520-4995
- OpenAlex
- W2017758139
- Cited by
- 112
- References
- 18
- Citations by year
- 2025: 3 · 2024: 2 · 2023: 1 · 2022: 2 · 2021: 5 · 2020: 3 · 2019: 8 · 2018: 3 · 2017: 6 · 2016: 2 · 2015: 4 · 2014: 6 · 2013: 6 · 2012: 4
- FWCI
- 3.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Redox biology and oxidative stress
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cell death mechanisms and regulation
- Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
- Keywords
- cytochrome c, tyrosine nitration, peroxynitrite, peroxidase activity, tetranitromethane, reactive nitrogen species, mitochondrial respiration
- MeSH terms
- Amino Acid Sequence, Animals, Electron Transport, Horses, Mitochondria, Heart, Molecular Sequence Data, Peroxidase, Potassium Cyanide, Sodium Cyanide, Tetranitromethane, Time Factors, Tyrosine, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Peroxynitrous Acid, Cytochromes c, Rats
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