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Abnormal tricarboxylic acid cycle metabolites in isovaleric acidaemia

  • Original Article
  • Published:
Journal of Inherited Metabolic Disease

Summary

Background

Although a number of abnormal diagnostic metabolites have previously been described in the urine of patients with isovaleric acidaemia (IVA), they do not fully explain the clinical symptoms associated with this disease.

Methods

On the basis of our current understanding of the TCA cycle and IVA, we predicted a number of abnormal methylated TCA cycle metabolites, initiated by methylsuccinic acid. We subsequently obtained characteristic gas chromatography–mass spectrometry elution times and mass spectra of the chemically synthesized predicted compounds and screened the urine of 6 IVA patients and 24 age-matched controls. Further proof for our findings was generated from a series of in vitro enzyme reactions using the chemically synthesized standards as substrates to their respective TCA cycle enzymes.

Results

Apart from the previously described methylsuccinic and methylfumaric acid, 3-methylmalic acid, (2R,3S)- and (2R,3R)-methylcitric acid and 2-methyl-cis-aconitic acid were detected in the urine of all 6 IVA patients in increased amounts. Additionally, although not directly determined, the in vitro enzyme reaction using of 3-methylmalic acid and malate dehydrogenase, in conjunction with the detection of 2-ketobutyric acid in the urine of all 6 IVA patients, strongly suggests an additional synthesis of 3-methyloxaloacetic acid by the same cycle.

Conclusion

Not only do these newly identified metabolites serve as additional diagnostic markers to those previously identified in IVA, but due to the structural arrangements of the (2R,3R)-methylcitric acid and 2-methyl-cis-aconitinic acid-derived 2-methylisocitric acid, inhibition of normal TCA cycle metabolism results at citrate synthase and isocitrate dehydrogenase, respectively.

Synopsis

Methylsuccinic acid acts as the initiating substrate to a series of abnormal, potentially harmful, methylated tricarboxylic acid cycle metabolites in isovaleric acidaemia.

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Abbreviations

GC-MS:

gas chromatography–mass spectrometry

IVA:

isovaleric acidaemia

TCA:

tricarboxylic acid cycle

References

  • Beach RL, Aogaichi T, Plaut GW (1977) Identification of d-threo-alpha-methylisocitrate as stereochemically specific substrate for bovine heart aconitase and inhibitor of TPN-linked isocitrate dehydrogenase. J Biol Chem 252: 2702–2709.

    PubMed  CAS  Google Scholar 

  • Budd MA, Tanaka K, Holmes LB, Efron ML, Crawford JD, Isselbacher KJ (1967) Isovaleric acidemia. Clinical features of a new genetic deect of leucine metabolism. N Engl J Med 277: 321–327.

    PubMed  CAS  Google Scholar 

  • Chalmers RA, Roe CR, Stacey TE, Hoppel CL (1984) Urinary excretion of l-carnitine and acylcarnitines by patients with disorders of organic acid metabolism: evidence for secondary insufficiency of l-carnitine. Pediatr Res 18: 1325–1328. doi:10.1203/00006450-198412000-00021.

    Article  PubMed  CAS  Google Scholar 

  • Coude FX, Ogier H, Grimber G, et al (1982) Correlation between blood ammonia concentration and organic acid accumulation in isovaleric and propionic acidemia. Pediatrics 69: 115–117.

    PubMed  CAS  Google Scholar 

  • Cox RFM, McElvain SM (1937) Ethyl ethoxyl propionate. Organic Synthesis 17: 272.

    Google Scholar 

  • Dorland L, Duran M, Wadman SK, Niederwieser A, Bruinvis L, Ketting D (1983) Isovalerylglucuronide, a new urinary metabolite in isovaleric acidemia. Identification problems due to rearrangement reactions. Clin Chim Acta 134: 77–83. doi:10.1016/0009-8981(83)90186-9.

    Article  PubMed  CAS  Google Scholar 

  • Gawron O, Mahajan KP (1966) Alpha-methyl-cis-aconitic acid, cis-aconitase substrate. I. Synthesis. Biochemistry 5: 2335–2342. doi:10.1021/bi00871a023.

    Article  PubMed  CAS  Google Scholar 

  • Kamerling JP, Duran M, Gerwig GJ, et al (1981) Determination of the absolute configuration of some biologically important urinary 2-hydroxydicarboxylic acids by capillary gas–liquid chromatography. J Chromatogr 222: 276–283.

    Article  PubMed  CAS  Google Scholar 

  • Lehnert W (1981a) 3-Hydroxyisoheptanoic acid: a new metabolite in isovaleric acidemia. Clin Chim Acta 113: 101–103. doi:10.1016/0009-8981(81)90445-9.

    Article  PubMed  CAS  Google Scholar 

  • Lehnert W (1981b) Excretion of N-isovalerylglutamic acid in isovaleric acidemia. Clin Chim Acta 116: 249–252. doi:10.1016/0009-8981(81)90030-9.

    Article  PubMed  CAS  Google Scholar 

  • Lehnert W (1983) N-Isovalerylalanine and N-isovalerylsarcosine: two new minor metabolites in isovaleric acidemia. Clin Chim Acta 134: 207–212. doi:10.1016/0009-8981(83)90198-5.

    Article  PubMed  CAS  Google Scholar 

  • Lehnert W, Niederhoff H (1981a) 4-hydroxyisovaleric acid: a new metabolite in isovaleric acidemia. Eur J Pediatr 136: 281–283. doi:10.1007/BF00442995.

    Article  PubMed  CAS  Google Scholar 

  • Lehnert W, Niederhoff H (1981b) 4-hydroxyisovaleric acid: a new metabolite in isovaleric acidemia. Eur J Pediatr 136: 281–283. doi:10.1007/BF00442995.

    Article  PubMed  CAS  Google Scholar 

  • Loots DT, Erasmus E, Mienie LJ (2005a) Identification of 19 new metabolites induced by abnormal amino acid conjugation in isovaleric acidemia. Clin Chem 51: 1510–1512. doi:10.1373/clinchem.2005.048421.

    Article  PubMed  CAS  Google Scholar 

  • Loots DT, Wiid IJ, Page BJ, Mienie LJ, van Helden PD (2005b) Melatonin prevents the free radical and MADD metabolic profiles induced by antituberculosis drugs in an animal model. J Pineal Res 38: 100–106. doi:10.1111/j.1600-079X.2004.00176.x.

    Article  PubMed  CAS  Google Scholar 

  • Nakamura E, Rosenberg LE, Tanaka K (1976) Microdetermination of methylmalonic acid and other short chain dicarboxylic acids by gas chromatography: use in prenatal diagnosis of methylmalonic acidemia and in studies of isovaleric acidemia. Clin Chim Acta 68: 127–140. doi:10.1016/0009-8981(76)90412-5.

    Article  PubMed  CAS  Google Scholar 

  • Nasser I, Mohsen AW, Jelesarov I, Vockley J, Macheroux P, Ghisla S (2004) Thermal unfolding of medium-chain acyl-CoA dehydrogenase and iso(3)valeryl-CoA dehydrogenase: study of the effect of genetic defects on enzyme stability. Biochim Biophys Acta 1690: 22–32.

    PubMed  CAS  Google Scholar 

  • Naylor G, Sweetman L, Nyhan WL, et al (1980) Isotope dilution analysis of methylcitric acid in amniotic fluid for the prenatal diagnosis of propionic and methylmalonic acidemia. Clin Chim Acta 107: 175–183. doi:10.1016/0009-8981(80)90445-3.

    Article  PubMed  CAS  Google Scholar 

  • Reformatskii S (1887) Neue Synthese zweiatomiger einbasischer Säuren aus den Ketonen. Chemische Berichte 20: 1210.

    Google Scholar 

  • Sweetman L, Williams JD (2001) Branched chain organic acidurias. In: Scriver CR, Beaudet AL, Sly WS, Valle D, eds; Childs B, Kinzler KW, Vogelstein B, assoc. eds. The Metabolic and Molecular Bases of Inherited Disease, 8th edn. New York: McGraw-Hill, 2125–2164.

    Google Scholar 

  • Tanaka K, Isselbacher KJ (1967) The isolation and identification of N-isovalerylglycine from urine of patients with isovaleric acidemia. J Biol Chem 242: 2966–2972.

    PubMed  CAS  Google Scholar 

  • Tanaka K, Rosenberg LE (1982) Disorders of branched chain amino acid and organic acid metabolism. In: Stanbury JB, Wyngaarden JB, Fredrickson DS, Goldstein JL, Brown MS, eds. The Metabolic Basis of Inherited Diseases. New York: McGraw-Hill, 440–473.

    Google Scholar 

  • Tanaka K, Orr JC, Isselbacher KJ (1968) Identification of beta-hydroxyisovaleric acid in the urine of a patient with isovaleric acidemia. Biochim Biophys Acta 152: 638–641.

    PubMed  CAS  Google Scholar 

  • Truscott RJ, Malegan D, McCairns E, et al (1981) New metabolites in isovaleric acidemia. Clin Chim Acta 110: 187–203. doi:10.1016/0009-8981(81)90348-X.

    Article  PubMed  CAS  Google Scholar 

  • van Rooyen JP, Mienie LJ, Erasmus E, de Wet WJ, Duran M, Wadman SK (1994) Urinary excretion of homocitric acid and methylhomocitric acid in propionic acidaemia: minor metabolic products of the citrate synthase aldol condensation reaction. Clin Chim Acta 230: 91–99. doi:10.1016/0009-8981(94)90092-2.

    Article  PubMed  Google Scholar 

  • Vockley J, Ensenauer R (2006). Isovaleric acidemia: new aspects of genetic and phenotypic heterogeneity. Am J Med Genet C Semin Med Genet 142C: 95–103. doi:10.1002/ajmg.c.30089.

    Article  PubMed  CAS  Google Scholar 

  • Vockley J, Parimoo B, Tanaka K (1991) Molecular characterization of four different classes of mutations in the isovaleryl-CoA dehydrogenase gene responsible for isovaleric acidemia. Am J Hum Genet 49: 147–157.

    PubMed  CAS  Google Scholar 

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Acknowledgements

I would like to thank the team at the North-West University Laboratory of Inborn Metabolic Defects for their assistance in this study.

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Authors and Affiliations

  1. School for Physical and Chemical Sciences, Centre of Metabolomics, Division of Biochemistry, North-West University, Hoffman Street, Private Bag X6001, Box 269, Potchefstroom, South Africa, 2531

    D. T. Loots

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  1. D. T. Loots
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Correspondence to D. T. Loots.

Additional information

Communicating editor: Moacir Wajner

Competing interests: None declared

References to electronic databases: Isovaleric acidaemia: OMIM #243500. Isovaleryl-CoA dehydrogenase: EC 1.3.99.10.

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Loots, D.T. Abnormal tricarboxylic acid cycle metabolites in isovaleric acidaemia. J Inherit Metab Dis 32, 403–411 (2009). https://doi.org/10.1007/s10545-009-1071-6

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  • DOI: https://doi.org/10.1007/s10545-009-1071-6

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