Human 2-Oxoglutarate Dehydrogenase and 2-Oxoadipate Dehydrogenase Both Generate Superoxide/H2O2 in a Side Reaction and Each Could Contribute to Oxidative Stress in Mitochondria

  • Frank JordanEmail author
  • Natalia NemeriaEmail author
  • Gary Gerfen
Original Paper


According to recent findings, the human 2-oxoglutarate dehydrogenase complex (hOGDHc) could be an important source of the reactive oxygen species in the mitochondria and could contribute to mitochondrial abnormalities associated with multiple neurodegenerative diseases, including Alzheimer’s disease, Huntington disease, and Parkinson’s disease. The human 2-oxoadipate dehydrogenase (hE1a) is a novel protein, which is encoded by the DHTKD1 gene. Both missence and nonsense mutations were identified in the DHTKD1 that lead to alpha-aminoadipic and alpha-oxoadipic aciduria, a metabolic disorder with a wide variety of the neurological abnormalities, and Charcot-Marie-Tooth disease type 2Q, an inherited neurological disorder affecting the peripheral nervous system. Recently, the rare pathogenic mutations in DHTKD1 and an increased H2O2 production were linked to the genetic ethiology of Eosinophilic Esophagitis (EoE), a chronic allergic inflammatory esophageal disorder. In view of the importance of hOGDHc in the tricarboxylic acid cycle (TCA cycle) and hE1a on the l-lysine, l-hydroxylysine and l-tryptophan degradation pathway in mitochondria, and to enhance our current understanding of the mechanism of superoxide/H2O2 generation by hOGDHc, and by human 2-oxoadipate dehydrogenase complex (hOADHc), this review focuses on several novel and unanticipated recent findings in vitro that emerged from the Jordan group’s research. Most significantly, the hE1o and hE1a now join the hE3 as being able to generate the superoxide/H2O2 in mitochondria.


2-Oxoglutarate and 2-oxoadipate dehydrogenase complexes Thiamin diphosphate-enamine radical Hydrogen peroxide Oxidative stress 



Human 2-oxoglutarate dehydrogenase complex


2-Oxoglutarate dehydrogenase, the first E1 component of hOGDHc;


Dihydrolipoyl succinyltransferase, the second E2 component of hOGDHc


Dihydrolipoyl dehydrogenase, the third E3 component of all 2-oxoacid dehydrogenase complexes


human 2-oxoadipate dehydrogenase complex, assembled from hE1a + hE2o + hE3


2-Oxoadipate dehydrogenase, the first component of hOADHc


Gene coding hE1a

TCA cycle

Tricarboxylic acid cycle


Hydrogen peroxide


Reactive oxygen species


Thiamin diphosphate








Electron Paramagnetic Resonance.



This work was supported, in whole or in part, by National Institutes of Health [Grant # 9R15GM116077-01 (to F.J.)]; the National Science Foundation [Grant CHE-1402675 (to F.J.) and Grant CHE 1213550 (to G.J.G)]; the Rutgers-Newark Chancellor’s SEED Grant (to F.J).

Compliance with Ethical Standards

Conflict of interest

The authors have no conflict of interest to declare.


  1. 1.
    Nemeria NS, Chakraborty S, Baykal A, Korotchkina LG, Patel MS, Jordan F (2007) The 1′,4′-iminopyrimidine tautomer of thiamin diphosphate is poised for catalysis in asymmetric active centers on enzymes. Proc Natl Acad Sci USA 104:78–82CrossRefPubMedGoogle Scholar
  2. 2.
    Nemeria NS, Chakraborty S, Balakrishnan A, Jordan F (2009) Reaction mechanisms of thiamin diphosphate enzymes:defining states of ionization and tautomerization of the cofactor at individual steps. FEBS J 276:2432–2446CrossRefPubMedPubMedCentralGoogle Scholar
  3. 3.
    Balakrishnan A, Gao Y, Moorjani P, Nemeria NS, Tittmann K, Jordan F (2012) Bifunctionality of the thiamin diphosphate cofactor: assignment of tautomeric/ionization states of the 4′-aminopyrimidine ring when various intermdediates occupy the active sites during the catalysis of yeast pyruvate decarboxylase. J Am Chem Soc 134:3873–3885CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Patel H, Nemeria NS, Brammer LA, Freel Meyers CL, Jordan F (2012) Observation of thiamin-bound intermediates and microscopic rate constants for their interconversion on 1-deoxy-d-xylulose 5-phosphate synthase: 600-fold rate acceleration of pyruvate decarboxylation by D-glyceraldehyde-3-phosphate. J Am Chem Soc 134:18374–18379CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Nemeria N, Binshtein E, Patel H, Balakrishnan A, Vered I, Shaanan B, Barak Z, Chipman D, Jordan F (2012) Glyoxylate carboligase: a unique thiamin diphosphate-dependent enzyme that can cycle between the 4′-aminopyrymidinium and 1′4′-iminopyrimidine tautomeric forms in the absence of the conserved glutamate. Biochemistry 51:7940–7952CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Jordan F, Patel H (2013) Catalysis in enzymatic decarboxylations: comparison of selected cofactor-dependent and cofactor-independent examples. ACS Catal 3:1601–1617CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Balakrishnan A, Jordan F, Nathan CF (2013) Influence of allosteric regulators on individual steps in the reaction catalyzed by Micobacterium tuberculosis 2-hydroxy-3-oxoadipate synthase. J Biol Chem 288:21688–21702CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Jordan F, Nemeria NS (2014) Progress in the experimental observation of thiamin diphosphate-bound intermediates on enzymes and mechanistic information derived from these observations. Bioorg Chem 57:251–262CrossRefPubMedGoogle Scholar
  9. 9.
    Nemeria NS, Ambrus A, Patel H, Gerfen G, Adam-Vizi V, Tretter L, Zhou J, Wang J, Jordan F (2014) Human 2-oxoglutarate dehydrogenase complex E1 component forms a thiamin-derived radical by aerobic oxidation of the enamine intermediate. J Biol Chem 289:29859–29873CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Nemeria NS, Gerfen G, Guevara E, Nareddy PR, Szostak M, Jordan F (2017) The human Krebs cycle 2-oxoglutarate dehydrogenase complex creates an additional source of superoxide/hydrogen peroxide from 2-oxoadipate as an alternative substrate. Free Radic Biol Med 108:644–654CrossRefPubMedGoogle Scholar
  11. 11.
    Zhou J, Yang L, Ozohanics O, Zhang X, Wang J, Ambrus A, Arjunan P, Brukh R, Nemeria NS, Furey W, Jordan F (2018) A multipronged approach unravels unprecedented protein- protein interactions in the human 2-oxoglutarate dehydrogenase multienzyme complex. J Biol Chem 293:19213–19227CrossRefPubMedGoogle Scholar
  12. 12.
    Nemeria NS, Gerfen G, Yang L, Zhang X, Jordan F (2018) Evidence for functional and regulatory cross-talk between the tricarboxylic acid cycle 2-oxoglutarate dehydrogenase complex and 2-oxoadipate dehydrogenase on the l-lysine, l-hydroxylysine and l-tryptophan degradation pathways from studies in vitro. Biochim Biophys Acta 1859:932–939. CrossRefGoogle Scholar
  13. 13.
    Nemeria NS, Gerfen G, Reddy Nareddy P, Yang L, Zhang X, Szostak M, Jordan F (2018) The mitochondrial 2-oxoadipate and 2-oxoglutarate dehydrogenase complexes share their E2 and E3 components for their function and both generate reactive oxygen species. Free Radic Biol Chem 115:136–145CrossRefGoogle Scholar
  14. 14.
    Chen H, Denton TT, Xu H, Calingasan N, Beal MF, Gibson GE (2016) Reductions in the mitochondrial enzyme α-ketoglutarate dehydrogenase complex in neurodegenerative disease-beneficial or detrimental? J Neurochem 139:823–839. CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Bunik VI, Sievers C (2002) Inactivation of the 2-oxo acid dehydrogenase complexes upon generation of intrinsic radical species. Eur J Biochem 269:5004–5015CrossRefPubMedGoogle Scholar
  16. 16.
    Starkov AA, Fiskum G, Chinopoulos C, Lorenzo BJ, Browne SE, Patel MS, Beal MF (2004) Mitochondrial alpha-ketoglutarate dehydrogenase complex generates reactive oxygen species. J Neurosci 24:7779–7788CrossRefPubMedGoogle Scholar
  17. 17.
    Tretter L, Adam-Vizi V (2004) Generation of reactive oxygen species in the reaction catalyzed by α-ketoglutarate dehydrogenase. J Neurosci 24:7771–7778CrossRefPubMedGoogle Scholar
  18. 18.
    Tretter L, Adam-Vizi V (2005) Alpha-ketoglutarate dehydrogenase: a target and generator of oxidative stress. Philos Trans R Soc Lond B Biol Sci 360:2335–2345CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Zündorf G, Kahlert S, Bunik VI, Reiser G (2009) α-Ketoglutarate dehydrogenase contributes to production of reactive oxygen species in glutamate-stimulated hippocampal neurons in situ. Neuroscience 158:610–616CrossRefPubMedGoogle Scholar
  20. 20.
    Quinlan CL, Goncalves RLS, Hey-Mogensen M, Yadava N, Bunik VI, Brand M (2014) The 2-oxoacid dehydrogenase complexes in mitochondria can produce superoxide/hydrogen peroxide at much higher rates than complex I. J Biol Chem 289:8312–8325CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Brand MD (2016) Mitochondrial generation of superoxide and hydrogen peroxide as the source of mitochondrial redox signaling. Free Rad Biol Med 100:14–31CrossRefPubMedGoogle Scholar
  22. 22.
    Danhauser K, Sauer SW, Haack TB, Wieland T, Staufner C, Graf E et al (2012) DHTKD1 mutations cause 2-aminoadipic and 2-oxoadipic aciduria. Am J Hum Genet 91:1082–1087CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Hagen J, Brinke H, Wanders RJA, Knegt AC, Oussoren E, Hoogeboom JM, Ruijter GJG, Becker D, Schwab KO, Franke I, Duran M, Waterham HR, Sass JO, Houten SM (2015) Genetic basis of alpha-aminoadipic and alpha-ketoadipic aciduria. J Inherit Metab Dis 38:873–879CrossRefPubMedGoogle Scholar
  24. 24.
    Stiles AR, Venturoni L, Mucci G, Elbalalesy N, Woontner M, Goodmann S, Abdenur JE (2015) New cases of DHTKD1 mutations in patients with 2-ketoadipic aciduria. JIMD Reports 25:15–19CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Xu WY, Gu MM, Sun LH, Guo WT, Zhu HB, Ma JF, Yuan WT, Kuang Y, Ji BJ, Wu XL, Chen Y, Zhang HX, Sun FT, Huang W, Huang L, Chen SD, Wang ZG (2012) A nonsence mutation in DHTKD1 causes Charcot-Marie-Tooth type 2 in a large chinese pedigree. Am J Hum Genet 91:1088–1094CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Xu W, Zhu H, Gu M, Luo Q, Ding J, Yao Y, Chen F, Wang Z (2013) DHTKD1 is essential for mitochondrial biogenesis and function maintenance. FEBS Lett 587:3587–3592CrossRefPubMedGoogle Scholar
  27. 27.
    Baets J, De Jonghe P, TimmermanV (2014) Recent advances in Charcot-Marie-Tooth disease. Curr Opin Neurol 27:532–540CrossRefPubMedGoogle Scholar
  28. 28.
    Xu WY, Zhu H, Shen Y, Wan YH, Tu XD, Wu WT, Tang L, Zhang HX, Lu SY, Jin XL, Fei J, Wang ZG (2018) DHTKD1 deficiency causes Charcot-Marie-Tooth disease in mice. Mol Cell Biol 38:e00085–e00018. PubMedPubMedCentralGoogle Scholar
  29. 29.
    Sherrill JD, Kc K, Wang X, Wen T, Chamberlin A, Stucke EM, Collins MH, Abonia JP, Peng Y, Wu Q, Putnam PE, Dexheimer PJ, Aronow BJ, Kottyan LC, Kaufman KM, Harley JB, Huang T, Rothenberg ME (2018) Whole-exome sequencing uncovers oxidoreductases DHTKD1 and OGDHc as linkers between mitochondrial disfynction and eosinophilic esophagitis. JCI Insight 3:1–20. CrossRefGoogle Scholar
  30. 30.
    Biagosch C, Ediga RD, Hensler SV, Faerberboeck M, Kuehn R, Wurst W, Meitinger T, Kölker S, Sauer S, Prokisch H (2017) Elevated glutaric acid levels in Dhtkd1-/Gcdh-double knockout mice challenge our current understanding of lysine metabolism. Biochim Biophys Acta - Molecular Basis of Disease 1863:2220–2228CrossRefPubMedGoogle Scholar
  31. 31.
    Schmiesing J, Lohmöller B, Schweizer M, Tidov H, Gersting SW, Muntau AG, Braulke T, Mühlhausen G (2017) Disease-causing mutations affecting surface residues of mitochondrial glutaryl-CoA dehydrogenase impair stability, heteromeric complex formation and mitochondria architecture. Hum Mol Genet 26:538–551PubMedGoogle Scholar
  32. 32.
    Goodman SI, Frerman FE (2001) Organic acidemias due to defects in lysine oxidation: 2-ketoadipic acidemia and glutaric acidemia. In: Scriver CR, Beaudet AL, Sly WS, Valle D, Childs B, Kinzler KW, Vogelstein B (eds) The metabolic and molecular basis of inherited disease, 8th edn. McGraw-Hill, pp 2195–2204Google Scholar
  33. 33.
    Tan M, Peng C, Anderson KA, Chhoy P, Xie Z, Dai L, Park J, Chen Y, Huang H, Zhang Y et al (2014)Lysine glutarylation is a protein pasttranslational modification regulated by SIRT5. Cell Metab 19:605–617Google Scholar
  34. 34.
    Schmiesing J, Storch S, Dörfler AC, Schweizer M, Makrypidi-Fraune G, Thelen M, Sylvester M, Gieselmann V, Meyer-Schwesinger C, Koch-Nolte F, Tidow H, Mühlhausen C, Waheed A, Sly WS, Braulke T (2018) Disease-linked glutarylation impairs function and interactions of mitochondrial proteins and contributes to mitochondrial heterogeneity. Cell Rep 24:2946–2956.
  35. 35.
    Reed LJ (2001) A trial of research from lipoic acid to α-keto acid dehydrogenase complexes. J Biol Chem 276:38329–38336CrossRefPubMedGoogle Scholar
  36. 36.
    Perham RN (1991) Domains, motifs, and linkers in 2-oxo acid dehydrogenase multienzyme complexes: a paradigm in the design of a multifunctional protein. Biochemistry 30:8501–8512CrossRefPubMedGoogle Scholar
  37. 37.
    Perham RN (2000) Swinging arms and swinging domains in multifunctional enzymes: catalytic machines for multistep reactions. Annu Rev Biochem 69:961–1004CrossRefPubMedGoogle Scholar
  38. 38.
    Yeaman SJ (1989) The 2-oxo acid dehydrogenase complexes: recent advances. Biochem J 257:625–632CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Maas E, Bisswanger H (1990) Localization of the alpha-oxoacid dehydrogenase multienzyme complexes within the mitochondrion. FEBS Lett 277:189–190CrossRefPubMedGoogle Scholar
  40. 40.
    Bunik VI, Tylicki A, Lukashev NY (2013) Thiamin diphosphate-dependent enzymes: from enzymology to metabolic regulation, drug designand disease models. FEBS J 280:6412–6442CrossRefPubMedGoogle Scholar
  41. 41.
    Frank RAW, Kay CWM, Hirst J, Luisi BF (2008) Off-Pathway, oxigen-dependent thiamine radical in krebs cycle. J Am Chem Soc 130:1662–1668CrossRefPubMedGoogle Scholar
  42. 42.
    Mansoorabadi SO, Seravalli J, Furdui C, Krymov V, Gerfen GJ, Begley TP, Melnick J, Ragsdale SW, Reed GH et al (2006) EPR spectroscopic and computational characterization of the hydroxyethylidene-thiamine pyrophosphate radical intermediate of pyruvate: ferredoxin oxidoreductase. Biochemistry 45:7122–7131CrossRefPubMedPubMedCentralGoogle Scholar
  43. 43.
    Ambrus A, Nemeria NS, Torocsik B, Tretter L, Nilsson M, Jordan F, Adam-Vizi V (2015) Formation of reactive oxygen species by human and bacterial pyruvate and 2-oxoglutarate dehydrogenase multienzyme complexes reconstituted from recombinant components. Free Radic Biol Med 89:642–650CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    Quinlan CL, Perevoschikova IV, Goncalves RL, Hey-Mogensen M, Brand MD (2013) The determination and analysis of site-specific rates of mitochondrial reactive oxygen species production. Methods Enzymol 526:189–217CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Goncalves RLS, Bunik VI, Brand MD (2016) Production of superoxide/hydrogen peroxide by the mitochondrial 2-oxoadipate dehydrogenase complex. Free Radic Biol Med 91:247–255CrossRefPubMedGoogle Scholar
  46. 46.
    Bunik VI, Brand MD (2018) Generation of superoxide and hydrogen peroxide by side reactions of mitochondrial 2-oxoacid dehydrogenase complexes in isolation and in cells. Biol Chem 399:407–420CrossRefPubMedGoogle Scholar
  47. 47.
    Hoffmann GF, Zschocke J (1999) Glutaric aciduria type I: from clinical, biochemical and molecular diversity to successful therapy. J Inherit Metab Dis 22:381–389CrossRefPubMedGoogle Scholar
  48. 48.
    Gibson GE, Xu H, Chen HL, Chen W, Denton T, Zhang S (2015) Alpha-ketoglutarate dehydrogenase complex-dependent succinylation of proteins in neurons and neuronal cell lines. J Neurochem 134:86–96CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Pan K, Jordan F (1998) DL-S-Methyllipoic acid methyl ester, a kinetically viable model for S-protonated lipoic acid as the oxidizing agent in reductive acyl transfers catalyzed by the 2-oxoacid dehydrogenase multienzyme complex. Biochemistry 37:1357–1364CrossRefPubMedGoogle Scholar

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Authors and Affiliations

  1. 1.Department of ChemistryRutgers UniversityNewarkUSA
  2. 2.Department of Physiology and BiophysicsAlbert Einstein College of MedicineBronxUSA

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