Cytochrome c reacts with cholesterol hydroperoxides to produce lipid- and protein-derived radicals

Thiago Cardoso Genaro De Mattos, Raphael F. Queiroz, Daniela Cunha, Patricia P. Appolinario, Paolo Di Mascio, Iseli L. Nantes, Ohara Augusto, Sayuri Miyamoto

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Lipid peroxidation is a well-known process that has been implicated in many diseases. Recent evidence has shown that mitochondrial cholesterol levels are increased under specific conditions, making it an important target for peroxidation inside the mitochondria. Cholesterol peroxidation generates, as primary products, several hydroperoxides (ChOOH), which can react with transition metals and metalloproteins. In this sense, cytochrome c (CYTC), a heme protein largely found in the mitochondria, becomes a candidate to react with ChOOH. Using CYTC associated with SDS micelles to mimic mitochondrial conditions, we show that ChOOH induces dose-dependent CYTC Soret band bleaching, indicating that it is using ChOOH as a substrate. This reaction leads to protein oligomerization, suggesting the formation of a protein radical that, subsequently, recombines, giving dimers, trimers, and tetramers. EPR experiments confirmed the production of carbon-centered radicals from both protein and lipid in the presence of ChOOH. Similar results were obtained with linoleic acid hydroperoxides (LAOOH). In addition, replacing SDS micelles by cardiolipin-containing liposomes as the mitochondrial mimetic led to similar results with either ChOOH or LAOOH. Importantly, kinetic experiments show that CYTC bleaching is faster with ChOOH than with H2O2, suggesting that these hydroperoxides could be relevant substrates for CYTC peroxidase-like activity in biological media. Altogether, these results show that CYTC induces homolytic cleavage of lipid-derived hydroperoxides, producing lipid and protein radicals.

Original languageEnglish (US)
Pages (from-to)2841-2850
Number of pages10
JournalBiochemistry
Volume54
Issue number18
DOIs
StatePublished - May 12 2015
Externally publishedYes

Fingerprint

Cytochromes c
Lipids
Mitochondria
Micelles
Bleaching
Proteins
Hydrogen Peroxide
Cytochrome-c Peroxidase
Cholesterol
Metalloproteins
Hemeproteins
Oligomerization
Cardiolipins
Lipid Peroxides
Substrates
Liposomes
Dimers
Lipid Peroxidation
Transition metals
Paramagnetic resonance

ASJC Scopus subject areas

  • Biochemistry
  • Medicine(all)

Cite this

Cardoso Genaro De Mattos, T., Queiroz, R. F., Cunha, D., Appolinario, P. P., Di Mascio, P., Nantes, I. L., ... Miyamoto, S. (2015). Cytochrome c reacts with cholesterol hydroperoxides to produce lipid- and protein-derived radicals. Biochemistry, 54(18), 2841-2850. https://doi.org/10.1021/bi501409d

Cytochrome c reacts with cholesterol hydroperoxides to produce lipid- and protein-derived radicals. / Cardoso Genaro De Mattos, Thiago; Queiroz, Raphael F.; Cunha, Daniela; Appolinario, Patricia P.; Di Mascio, Paolo; Nantes, Iseli L.; Augusto, Ohara; Miyamoto, Sayuri.

In: Biochemistry, Vol. 54, No. 18, 12.05.2015, p. 2841-2850.

Research output: Contribution to journalArticle

Cardoso Genaro De Mattos, T, Queiroz, RF, Cunha, D, Appolinario, PP, Di Mascio, P, Nantes, IL, Augusto, O & Miyamoto, S 2015, 'Cytochrome c reacts with cholesterol hydroperoxides to produce lipid- and protein-derived radicals', Biochemistry, vol. 54, no. 18, pp. 2841-2850. https://doi.org/10.1021/bi501409d
Cardoso Genaro De Mattos, Thiago ; Queiroz, Raphael F. ; Cunha, Daniela ; Appolinario, Patricia P. ; Di Mascio, Paolo ; Nantes, Iseli L. ; Augusto, Ohara ; Miyamoto, Sayuri. / Cytochrome c reacts with cholesterol hydroperoxides to produce lipid- and protein-derived radicals. In: Biochemistry. 2015 ; Vol. 54, No. 18. pp. 2841-2850.
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