Mitochondrial α-ketoglutarate dehydrogenase complex generates reactive oxygen species

Anatoly A. Starkov, Gary Fiskum, C. Chinopoulos, Beverly J. Lorenzo, Susan E. Browne, Mulchand S. Patel, M. Flint Beal

Research output: Contribution to journalArticle

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Abstract

Mitochondria-produced reactive oxygen species (ROS) are thought to contribute to cell death caused by a multitude of pathological conditions. The molecular sites of mitochondrial ROS production are not well established but are generally thought to be located in complex I and complex III of the electron transport chain. We measured H2O2 production, respiration, and NADPH reduction level in rat brain mitochondria oxidizing a variety of respiratory substrates. Under conditions of maximum respiration induced with either ADP or carbonyl cyanide p-trifluoromethoxyphenylhydrazone, α-ketoglutarate supported the highest rate of H2O2 production. In the absence of ADP or in the presence of rotenone, H 2O2 production rates correlated with the reduction level of mitochondrial NADPH with various substrates, with the exception of α-ketoglutarate. Isolated mitochondrial α-ketoglutarate dehydrogenase (KGDHC) and pyruvate dehydrogenase (PDHC) complexes produced superoxide and H2O2. NAD+ inhibited ROS production by the isolated enzymes and by permeabilized mitochondria. We also measured H2O2 production by brain mitochondria isolated from heterozygous knock-out mice deficient in dihydrolipoyl dehydrogenase (Did). Although this enzyme is a part of both KGDHC and PDHC, there was greater impairment of KGDHC activity in Dld-deficient mitochondria. These mitochondria also produced significantly less H2O2 than mitochondria isolated from their littermate wild-type mice. The data strongly indicate that KGDHC is a primary site of ROS production in normally functioning mitochondria.

Original languageEnglish
Pages (from-to)7779-7788
Number of pages10
JournalJournal of Neuroscience
Volume24
Issue number36
DOIs
Publication statusPublished - Sep 8 2004

Fingerprint

Ketoglutarate Dehydrogenase Complex
Reactive Oxygen Species
Mitochondria
NADP
Adenosine Diphosphate
Respiration
Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
Dihydrolipoamide Dehydrogenase
Pyruvate Dehydrogenase Complex
Rotenone
Electron Transport Complex III
Brain
Enzymes
Knockout Mice
Superoxides
NAD
Oxidoreductases
Cell Death

Keywords

  • Alzheimer
  • Ketoglutarate dehydrogenase
  • Lipoamide dehydrogenase
  • Mitochondria
  • Parkinson
  • Reactive oxygen species

ASJC Scopus subject areas

  • Neuroscience(all)

Cite this

Starkov, A. A., Fiskum, G., Chinopoulos, C., Lorenzo, B. J., Browne, S. E., Patel, M. S., & Beal, M. F. (2004). Mitochondrial α-ketoglutarate dehydrogenase complex generates reactive oxygen species. Journal of Neuroscience, 24(36), 7779-7788. https://doi.org/10.1523/JNEUROSCI.1899-04.2004

Mitochondrial α-ketoglutarate dehydrogenase complex generates reactive oxygen species. / Starkov, Anatoly A.; Fiskum, Gary; Chinopoulos, C.; Lorenzo, Beverly J.; Browne, Susan E.; Patel, Mulchand S.; Beal, M. Flint.

In: Journal of Neuroscience, Vol. 24, No. 36, 08.09.2004, p. 7779-7788.

Research output: Contribution to journalArticle

Starkov, AA, Fiskum, G, Chinopoulos, C, Lorenzo, BJ, Browne, SE, Patel, MS & Beal, MF 2004, 'Mitochondrial α-ketoglutarate dehydrogenase complex generates reactive oxygen species', Journal of Neuroscience, vol. 24, no. 36, pp. 7779-7788. https://doi.org/10.1523/JNEUROSCI.1899-04.2004
Starkov, Anatoly A. ; Fiskum, Gary ; Chinopoulos, C. ; Lorenzo, Beverly J. ; Browne, Susan E. ; Patel, Mulchand S. ; Beal, M. Flint. / Mitochondrial α-ketoglutarate dehydrogenase complex generates reactive oxygen species. In: Journal of Neuroscience. 2004 ; Vol. 24, No. 36. pp. 7779-7788.
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