Home LiteratureArticle Details
PMID: 11124972 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Inhibition of Krebs cycle enzymes by hydrogen peroxide: A key role of [alpha]-ketoglutarate dehydrogenase in limiting NADH production under oxidative stress.

The Journal of neuroscience : the official journal of the Society for Neuroscience ·Vol. 20 ·No. 24 ·2000-12-15 ·Pages 8972-9

Tretter L, Adam-Vizi V

Abstract

In this study we addressed the function of the Krebs cycle to determine which enzyme(s) limits the availability of reduced nicotinamide adenine dinucleotide (NADH) for the respiratory chain under H(2)O(2)-induced oxidative stress, in intact isolated nerve terminals. The enzyme that was most vulnerable to inhibition by H(2)O(2) proved to be aconitase, being completely blocked at 50 microm H(2)O(2). alpha-Ketoglutarate dehydrogenase (alpha-KGDH) was also inhibited but only at higher H(2)O(2) concentrations (>/=100 microm), and only partial inactivation was achieved. The rotenone-induced increase in reduced nicotinamide adenine dinucleotide (phosphate) [NAD(P)H] fluorescence reflecting the amount of NADH available for the respiratory chain was also diminished by H(2)O(2), and the effect exerted at small concentrations (</=50 microm) of the oxidant was completely prevented by 1,3-bis(2-chloroethyl)-1-nitrosourea (BCNU), an inhibitor of glutathione reductase. BCNU-insensitive decline by H(2)O(2) in the rotenone-induced NAD(P)H fluorescence correlated with inhibition of alpha-ketoglutarate dehydrogenase. Decrease in the glutamate content of nerve terminals was induced by H(2)O(2) at concentrations inhibiting aconitase. It is concluded that (1) aconitase is the most sensitive enzyme in the Krebs cycle to inhibition by H(2)O(2), (2) at small H(2)O(2) concentrations (</=50 microm) when aconitase is inactivated, glutamate fuels the Krebs cycle and NADH generation is unaltered, (3) at higher H(2)O(2) concentrations (>/=100 microm) inhibition of alpha-ketoglutarate dehydrogenase limits the amount of NADH available for the respiratory chain, and (4) increased consumption of NADPH makes a contribution to the H(2)O(2)-induced decrease in the amount of reduced pyridine nucleotides. These results emphasize the importance of alpha-KGDH in impaired mitochondrial function under oxidative stress, with implications for neurodegenerative diseases and cell damage induced by ischemia/reperfusion.

MeSH Terms
Aconitate Hydratase/antagonists & inhibitors,metabolism Animals Carmustine/pharmacology Cerebral Cortex/chemistry,enzymology Citrate (si)-Synthase/metabolism,physiology Citric Acid Cycle/drug effects Dose-Response Relationship, Drug Electron Transport/drug effects Enzyme Activation/drug effects Glucose/metabolism Glutamic Acid/metabolism Glutathione Reductase/antagonists & inhibitors,metabolism Guinea Pigs Hydrogen Peroxide/pharmacology Ketoglutarate Dehydrogenase Complex/antagonists & inhibitors,metabolism Malate Dehydrogenase/metabolism NAD/biosynthesis NADP/metabolism Oxidative Stress/drug effects Pyrimidines/metabolism Rotenone/pharmacology Spectrometry, Fluorescence Succinate Dehydrogenase/metabolism Synaptosomes/chemistry,enzymology
Chemicals
Pyrimidines Rotenone NAD Glutamic Acid NADP Hydrogen Peroxide Malate Dehydrogenase Ketoglutarate Dehydrogenase Complex Succinate Dehydrogenase Glutathione Reductase Citrate (si)-Synthase Aconitate Hydratase Glucose Carmustine
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Tretter L
Department of Medical Biochemistry, Neurochemical Group, Semmelweis University of Medicine, Budapest, H-1444, Hungary.
Adam-Vizi V
References (58)
58 references, click to expand
  1. Active site-specific inhibition by 1,3-bis(2-chloroethyl)-1-nitrosourea of two genetically homologous flavoenzymes: glutathione reductase and lipoamide dehydrogenase.
    J Lab Clin Med. 1985 Apr;105(4):464-71 PMID: 3920338
  2. Metabolic and energetic properties of isolated nerve ending particles (synaptosomes).
    Biochim Biophys Acta. 1996 Nov 12;1277(1-2):13-34 PMID: 8950370
  3. A simple ultramicro method for determination of pyridine nucleotides in tissues.
    Anal Biochem. 1969 Feb;27(2):212-7 PMID: 4388021
  4. Isolated cerebral and cerebellar mitochondria produce free radicals when exposed to elevated CA2+ and Na+: implications for neurodegeneration.
    J Neurochem. 1994 Aug;63(2):584-91 PMID: 8035183
  5. Abnormalities of mitochondrial enzymes in Alzheimer disease.
    J Neural Transm (Vienna). 1998;105(8-9):855-70 PMID: 9869323
  6. A "radical" view of cerebral ischemic injury.
    Prog Neurobiol. 1994 Mar;42(4):441-8 PMID: 8090929
  7. Comparison of the structures of the quinone-binding sites in beef heart mitochondria.
    J Biol Chem. 1993 Sep 15;268(26):19328-33 PMID: 8396133
  8. Measuring nitric oxide and superoxide: rate constants for aconitase reactivity.
    Methods Enzymol. 1996;269:37-41 PMID: 8791635
  9. Declines in mitochondrial respiration during cardiac reperfusion: age-dependent inactivation of alpha-ketoglutarate dehydrogenase.
    Proc Natl Acad Sci U S A. 1999 Jun 8;96(12):6689-93 PMID: 10359773
  10. The oxidative inactivation of mitochondrial electron transport chain components and ATPase.
    J Biol Chem. 1990 Sep 25;265(27):16330-6 PMID: 2168888
  11. Inactivation-reactivation of aconitase in Escherichia coli. A sensitive measure of superoxide radical.
    J Biol Chem. 1992 May 5;267(13):8757-63 PMID: 1315737
  12. Identification of free hydroxyl radicals in respiring rat heart mitochondria by spin trapping with the nitrone DMPO.
    FEBS Lett. 1981 Jan 26;123(2):241-4 PMID: 6262109
  13. Glucose and synaptosomal glutamate metabolism: studies with [15N]glutamate.
    J Neurochem. 1988 Sep;51(3):892-902 PMID: 2900879
  14. The alpha-ketoglutarate dehydrogenase complex in neurodegeneration.
    Neurochem Int. 2000 Feb;36(2):97-112 PMID: 10676873
  15. Depolarization of in situ mitochondria due to hydrogen peroxide-induced oxidative stress in nerve terminals: inhibition of alpha-ketoglutarate dehydrogenase.
    J Neurochem. 1999 Jul;73(1):220-8 PMID: 10386974
  16. An ultrastructural analysis of the effects of accumulation of neurofibrillary tangle in pyramidal neurons of the cerebral cortex in Alzheimer's disease.
    Neuropathol Appl Neurobiol. 1986 May-Jun;12(3):305-19 PMID: 3736776
  17. Sequestration of glutamate-induced Ca2+ loads by mitochondria in cultured rat hippocampal neurons.
    J Neurophysiol. 1996 Sep;76(3):1611-21 PMID: 8890280
  18. Mechanisms of oxidant-mediated cell injury. The glycolytic and mitochondrial pathways of ADP phosphorylation are major intracellular targets inactivated by hydrogen peroxide.
    J Biol Chem. 1988 Feb 5;263(4):1665-75 PMID: 3338986
  19. Superoxide radical and iron modulate aconitase activity in mammalian cells.
    J Biol Chem. 1995 Jun 2;270(22):13399-405 PMID: 7768942
  20. Early events in free radical-mediated damage of isolated nerve terminals: effects of peroxides on membrane potential and intracellular Na+ and Ca2+ concentrations.
    J Neurochem. 1996 May;66(5):2057-66 PMID: 8780036
  21. Effect of succinate on mitochondrial lipid peroxidation. 2. The protective effect of succinate against functional and structural changes induced by lipid peroxidation.
    J Bioenerg Biomembr. 1987 Feb;19(1):31-44 PMID: 3032929
  22. Brain alpha-ketoglutarate dehydrogenase complex: kinetic properties, regional distribution, and effects of inhibitors.
    J Neurochem. 1986 Nov;47(5):1376-86 PMID: 3760866
  23. Peroxide modulation of slow onset potentiation in rat hippocampus.
    J Neurosci. 1997 Nov 15;17(22):8695-701 PMID: 9348338
  24. Hydroperoxide-induced oxidative stress alters pyridine nucleotide metabolism in neonatal heart muscle cells.
    Am J Physiol. 1993 Jun;264(6 Pt 1):C1401-10 PMID: 8392794
  25. Generation of hydrogen peroxide by brain mitochondria: the effect of reoxygenation following postdecapitative ischemia.
    Arch Biochem Biophys. 1989 Mar;269(2):623-38 PMID: 2919886
  26. Influence of the malate-aspartate shuttle on oxidative metabolism in synaptosomes.
    J Neurochem. 1988 May;50(5):1559-65 PMID: 3361310
  27. Enhanced depolarization-evoked calcium signal and reduced [ATP]/[ADP] ratio are unrelated events induced by oxidative stress in synaptosomes.
    J Neurochem. 1997 Dec;69(6):2529-37 PMID: 9375686
  28. Astrocytes protect neurons from hydrogen peroxide toxicity.
    J Neurosci. 1996 Apr 15;16(8):2553-62 PMID: 8786431
  29. An NADH-linked spectrophotometric assay for pyruvate dehydrogenase complex in crude tissue homogenates.
    J Biol Chem. 1981 Jul 10;256(13):6583-6 PMID: 7240230
  30. Exacerbated responses to oxidative stress by an Na(+) load in isolated nerve terminals: the role of ATP depletion and rise of [Ca(2+)](i).
    J Neurosci. 2000 Mar 15;20(6):2094-103 PMID: 10704483
  31. Inhibition of NADH-linked mitochondrial respiration by 4-hydroxy-2-nonenal.
    Biochemistry. 1998 Jan 13;37(2):552-7 PMID: 9425076
  32. The glutathione system of peroxide detoxification is less efficient in neurons than in astroglial cells.
    J Neurochem. 1999 Jun;72(6):2523-30 PMID: 10349863
  33. The regeneration of reduced glutathione in rat forebrain mitochondria identifies metabolic pathways providing the NADPH required.
    Neurosci Lett. 1999 Nov 12;275(2):97-100 PMID: 10568508
  34. Inactivation of aconitase and oxoglutarate dehydrogenase in skeletal muscle in vitro by superoxide anions and/or nitric oxide.
    Biochem Biophys Res Commun. 1998 Aug 19;249(2):512-6 PMID: 9712727
  35. Measurement of striatal H2O2 by microdialysis following global forebrain ischemia and reperfusion in the rat: correlation with the cytotoxic potential of H2O2 in vitro.
    Brain Res. 1995 Feb 13;671(2):181-6 PMID: 7743206
  36. Tricarboxylic acid cycle flux and enzyme activities in the isolated working rat heart.
    Biochem J. 1981 Dec 15;200(3):701-3 PMID: 7342978
  37. Mitochondrial depolarization in glutamate-stimulated neurons: an early signal specific to excitotoxin exposure.
    J Neurosci. 1996 Sep 15;16(18):5688-97 PMID: 8795624
  38. A radical hypothesis for neurodegeneration.
    Trends Neurosci. 1993 Nov;16(11):439-44 PMID: 7507613
  39. Glutamate, calcium, and free radicals as mediators of ischemic brain damage.
    Ann Thorac Surg. 1995 May;59(5):1316-20 PMID: 7733760
  40. Cytosolic Ca2+ changes during in vitro ischemia in rat hippocampal slices: major roles for glutamate and Na+-dependent Ca2+ release from mitochondria.
    J Neurosci. 1999 May 1;19(9):3307-15 PMID: 10212290
  41. Influence of NAD-linked dehydrogenase activity on flux through oxidative phosphorylation.
    Biochem J. 1990 Jun 1;268(2):421-8 PMID: 2363681
  42. Mitochondria, free radicals, and neurodegeneration.
    Curr Opin Neurobiol. 1996 Oct;6(5):661-6 PMID: 8937831
  43. The role of oxidative abnormalities in the pathophysiology of Alzheimer's disease.
    Rev Neurol (Paris). 1991;147(6-7):513-25 PMID: 1962057
  44. Tricarboxylic acid cycle in rat brain synaptosomes. Fluxes and interactions with aspartate aminotransferase and malate/aspartate shuttle.
    J Biol Chem. 1994 Nov 4;269(44):27414-20 PMID: 7961653
  45. Role of calcium ions in regulation of mammalian intramitochondrial metabolism.
    Physiol Rev. 1990 Apr;70(2):391-425 PMID: 2157230
  46. 1,3-Bis(2-chloroethyl)-1-nitrosourea as thiol-carbamoylating agent in biological systems.
    Methods Enzymol. 1995;251:173-88 PMID: 7651196
  47. Oxidative damage, mitochondrial oxidant generation and antioxidant defenses during aging and in response to food restriction in the mouse.
    Mech Ageing Dev. 1994 May;74(1-2):121-33 PMID: 7934203
  48. An immunohistochemical study on alpha-ketoglutarate dehydrogenase complex in Parkinson's disease.
    Ann Neurol. 1994 Feb;35(2):204-10 PMID: 8109900
  49. Alpha-ketoglutarate dehydrogenase in Alzheimer brains bearing the APP670/671 mutation.
    Ann Neurol. 1998 Oct;44(4):676-81 PMID: 9778267
  50. Superoxide radicals as precursors of mitochondrial hydrogen peroxide.
    FEBS Lett. 1974 May 15;42(1):68-72 PMID: 4859511
  51. Glutathione reductase.
    Methods Enzymol. 1985;113:484-90 PMID: 3003504
  52. An improved cycling assay for nicotinamide adenine dinucleotide.
    Anal Biochem. 1973 Jun;53(2):452-8 PMID: 4351948
  53. The cellular production of hydrogen peroxide.
    Biochem J. 1972 Jul;128(3):617-30 PMID: 4404507
  54. Metabolic impairment induces oxidative stress, compromises inflammatory responses, and inactivates a key mitochondrial enzyme in microglia.
    J Neurochem. 1999 May;72(5):1948-58 PMID: 10217272
  55. High-performance liquid chromatography analysis of oxidized and reduced pyridine dinucleotides in specific brain regions.
    Anal Biochem. 1995 Jul 1;228(2):312-7 PMID: 8572312
  56. Requirement for superoxide in excitotoxic cell death.
    Neuron. 1996 Feb;16(2):345-55 PMID: 8789949
  57. Apoptotic vs. nonapoptotic cytotoxicity induced by hydrogen peroxide.
    Free Radic Biol Med. 1997;22(1-2):73-83 PMID: 8958131
  58. Effects of oxidants and glutamate receptor activation on mitochondrial membrane potential in rat forebrain neurons.
    J Neurochem. 1998 Dec;71(6):2392-400 PMID: 9832137
Article Info
Journal
The Journal of neuroscience : the official journal of the Society for Neuroscience
Abbr.
J Neurosci
ISSN
1529-2401
Published
2000-12-15
Pages
8972-9
Language
English
Region
United States
NLM ID
8102140
PMCID
PMC6773008
Subset
IM
External Links
PubMed source PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2