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Lactate Dehydrogenase Activity is Inhibited by Methylmalonate in vitro

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Abstract

Methylmalonic acidemia (MMAemia) is an inherited metabolic disorder of branched amino acid and odd-chain fatty acid metabolism, involving a defect in the conversion of methylmalonyl-coenzyme A to succinyl-coenzyme A. Systemic and neurological manifestations in this disease are thought to be associated with the accumulation of methylmalonate (MMA) in tissues and biological fluids with consequent impairment of energy metabolism and oxidative stress. In the present work we studied the effect of MMA and two other inhibitors of mitochondrial respiratory chain complex II (malonate and 3-nitropropionate) on the activity of lactate dehydrogenase (LDH) in tissue homogenates from adult rats. MMA potently inhibited LDH-catalyzed conversion of lactate to pyruvate in liver and brain homogenates as well as in a purified bovine heart LDH preparation. LDH was about one order of magnitude less sensitive to inhibition by MMA when catalyzing the conversion of pyruvate to lactate. Kinetic studies on the inhibition of brain LDH indicated that MMA inhibits this enzyme competitively with lactate as a substrate (K i=3.02±0.59 mM). Malonate and 3-nitropropionate also strongly inhibited LDH-catalyzed conversion of lactate to pyruvate in brain homogenates, while no inhibition was observed by succinate or propionate, when present in concentrations of up to 25 mM. We propose that inhibition of the lactate/pyruvate conversion by MMA contributes to lactate accumulation in blood, metabolic acidemia and inhibition of gluconeogenesis observed in patients with MMAemia. Moreover, the inhibition of LDH in the central nervous system may also impair the lactate shuttle between astrocytes and neurons, compromising neuronal energy metabolism.

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References

  1. Coulombe JT, Shih VE, Levy HL (1981) Massachusetts metabolic disorders screening program. II. Methylmalonic aciduria. Pediatrics 67:26–31

    PubMed  CAS  Google Scholar 

  2. Fenton WA, Gravel RA, Rosenblatt DS (2001) Disorders of propionate and methylmalonate metabolism. In: Scriver CR, Beaudet AL, Valle AD, Sly W (eds) The metabolic and molecular basis of inherited disease, 8th edn. McGraw-Hill, New York, pp 2165–2193

    Google Scholar 

  3. Hoffmann GF, Gibson KM, Trefz FK, Nyhan WL, Bremer HJ, Rating D (1994) Neurological manifestations of organic acid disorders. Eur J Pediatr 153:S94–S100

    Article  PubMed  CAS  Google Scholar 

  4. Hoffmann GF, Meier-Augenstein W, Stöckler S, Surtees R, Rating D, Nyhan WL (1993) Physiology and pathophysiology of organic acids in cerebrospinal fluid. J Inherit Metab Dis 16:648–669

    Article  PubMed  CAS  Google Scholar 

  5. Leonard JV (1995) The management and outcome of propionic and methylmalonic acidaemia. J Inherit Metab Dis 18:430–434

    Article  PubMed  CAS  Google Scholar 

  6. de Baulny HO, Benoist JF, Rigal O, Touati G, Rabier D, Saudubray JM (2005) Methylmalonic and propionic acidaemias: management and outcome. J Inherit Metab Dis 28:415–423

    Article  PubMed  CAS  Google Scholar 

  7. Maciel EN, Kowaltowski AJ, Schwalm FD, Rodrigues JM, Souza DO, Vercesi AE, Wajner M, Castilho RF (2004) Mitochondrial permeability transition in neuronal damage promoted by Ca2+ and respiratory chain complex II inhibition. J Neurochem 90:1025–1035

    Article  PubMed  CAS  Google Scholar 

  8. Kowaltowski AJ, Castilho RF, Vercesi AE (2001) Mitochondrial permeability transition and oxidative stress. FEBS Lett 495:12–15

    Article  PubMed  CAS  Google Scholar 

  9. Green DR, Kroemer G (2004) The pathophysiology of mitochondrial cell death. Science 305:626–629

    Article  PubMed  CAS  Google Scholar 

  10. Heidenreich R, Natowicz M, Hainline BE, Berman P, Kelley RI, Hillman RE, Berry GT (1988) Acute extrapyramidal syndrome in methylmalonic acidemia: “metabolic stroke” involving the globus pallidus. J Pediatr 113:1022–1027

    Article  PubMed  CAS  Google Scholar 

  11. Trinh BC, Melhem ER, Barker PB (2001) Multi-slice proton MR spectroscopy and diffusion-weighted imaging in methylmalonic acidemia: report of two cases and review of the literature. Am J Neuroradiol 22:831–833

    PubMed  CAS  Google Scholar 

  12. Rosenberg LE (1978) Disorders of propionate, methylmalonate and cobalamin metabolism. In: Stanbury JB, Wyngaarden JB, Fredrickson DS (eds) The metabolic basis of inherited disease, 4th edn. McGraw-Hill, New York, p 411

    Google Scholar 

  13. Worthen HG, Al Ashwal A, Ozand PT, Garawi S, Rahbeeni Z, Al Odaib A, Subramanyam SB, Rashed M (1994) Comparative frequency and severity of hypoglycemia in selected organic acidemias, branched chain amino acidemia, and disorders of fructose metabolism. Brain Dev 16(Suppl):81–85

    Article  PubMed  Google Scholar 

  14. Oberholzer VG, Levin B, Burgess EA, Young WF (1967) Methylmalonic aciduria. An inborn error of metabolism leading to chronic metabolic acidosis. Arch Dis Child 42:492–504

    Article  PubMed  CAS  Google Scholar 

  15. Halperin ML, Schiller CM, Fritz IB (1971) The inhibition by methylmalonic acid of malate transport by the dicarboxylate carrier in rat liver mitochondria. A possible explanation for hypoglycemia in methylmalonic aciduria. J Clin Invest 50:2276–2282

    Article  PubMed  CAS  Google Scholar 

  16. Dutra JC, Dutra-Filho CS, Cardozo SE, Wannmacher CM, Sarkis JJ, Wajner M (1993) Inhibition of succinate dehydrogenase and beta-hydroxybutyrate dehydrogenase activities by methylmalonate in brain and liver of developing rats. J Inherit Metab Dis 16:147–153

    Article  PubMed  CAS  Google Scholar 

  17. Brusque AM, Borba Rosam R, Schuckm PF, Dalcin KB, Ribeiro CA, Silva CG, Wannmacher CM, Dutra-Filho CS, Wyse AT, Briones P, Wajner M (2002) Inhibition of the mitochondrial respiratory chain complex activities in rat cerebral cortex by methylmalonic acid. Neurochem Int 40:593–601

    Article  PubMed  CAS  Google Scholar 

  18. Okun JG, Horster F, Farkas LM, Feyh P, Hinz A, Sauer S, Hoffmann GF, Unsicker K, Mayatepek E, Kolker S (2002) Neurodegeneration in methylmalonic aciduria involves inhibition of complex II and the tricarboxylic acid cycle, and synergistically acting excitotoxicity. J Biol Chem 277:14674–14680

    PubMed  CAS  Google Scholar 

  19. Kolker S, Schwab M, Horster F, Sauer S, Hinz A, Wolf NI, Mayatepek E, Hoffmann GF, Smeitink JA, Okun JG (2003) Methylmalonic acid, a biochemical hallmark of methylmalonic acidurias but no inhibitor of mitochondrial respiratory chain. J Biol Chem 278:47388–47393

    Article  PubMed  Google Scholar 

  20. Schuck PF, Rosa RB, Pettenuzzo LF, Sitta A, Wannmacher CM, Wyse AT, Wajner M (2004) Inhibition of mitochondrial creatine kinase activity from rat cerebral cortex by methylmalonic acid. Neurochem Int 45:661–667

    Article  PubMed  CAS  Google Scholar 

  21. Gladden LB (2004) Lactate metabolism: a new paradigm for the third millennium. J Physiol 558:5–30

    Article  PubMed  CAS  Google Scholar 

  22. Gornall AG, Bardawill CJ, Donid MM (1949) Determination of serum protein by means of the Biuret reaction. J Biol Chem 177:751–766

    PubMed  CAS  Google Scholar 

  23. Sahlin K, Katz A, Broberg S (1990) Tricarboxylic acid cycle intermediates in human muscle during prolonged exercise. Am J Physiol 259:C834–841

    PubMed  CAS  Google Scholar 

  24. Spencer MK, Katz A, Raz I (1991) Epinephrine increases tricarboxylic acid cycle intermediates in human skeletal muscle. Am J Physiol 260:E436–E439

    PubMed  CAS  Google Scholar 

  25. Lehmann HP, Henry JB (2001) SI Units. In: Henry JB (ed) Clinical diagnosis and management by laboratory methods, 20th edn. W.B. Saunders Company, Philadelphia, USA, pp 1426–1441

    Google Scholar 

  26. Comar JF, Suzuki-Kemmelmeier F, Bracht A (2003) The action of oxybutynin on haemodynamics and metabolism in the perfused rat liver. Pharmacol Toxicol 93:147–152

    Article  PubMed  CAS  Google Scholar 

  27. Hakala MT, Glaid AJ, Schwert GW (1956) Lactic dehydrogenase. II. Variation of kinetic and equilibrium constants with temperature. J Biol Chem 221:191–209

    PubMed  CAS  Google Scholar 

  28. Dixon M, Webb EC (eds) (1964) Enzyme kinetics, 2nd edn. Longman, London, UK

    Google Scholar 

  29. Kaplan NO, Everse J (1972) Regulatory characteristics of lactate dehydrogenases. Adv Enzyme Regul 10:323–336

    Article  PubMed  CAS  Google Scholar 

  30. Bittar PG, Charnay Y, Pellerin L, Bouras C, Magistretti PJ (1996) Selective distribution of lactate dehydrogenase isoenzymes in neurons and astrocytes of human brain. J Cereb Blood Flow Metab 16:1079–1089

    Article  PubMed  CAS  Google Scholar 

  31. Brooks GA, Dubouchaud H, Brown M, Sicurello JP, Butz CE (1999) Role of mitochondrial lactate dehydrogenase and lactate oxidation in the intracellular lactate shuttle. Proc Natl Acad Sci USA 96:1129–1134

    Article  PubMed  CAS  Google Scholar 

  32. Wajner M, Coelho JC (1997) Neurological dysfunction in methylmalonic acidaemia is probably related to the inhibitory effect of methylmalonate on brain energy production. J Inherit Metab Dis 20:761–768

    Article  PubMed  CAS  Google Scholar 

  33. Pellerin L, Magistretti PJ (1994) Glutamate uptake into astrocytes stimulates aerobic glycolysis: a mechanism coupling neuronal activity to glucose utilization. Proc Natl Acad Sci USA 91:10625–10629

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We thank Dr. Alicia Kowaltowski for discussions and Edilene S. Santos and Elisangela J. Gomes for technical support. This study was supported by Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) and National Institutes of Health (NIH) (grant number: 1 R03 HD047388–01A1). L.O.S and S.R.M. were supported by CNPq fellowships and E.N.M. by a FAPESP fellowship.

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Authors and Affiliations

  1. Departamento de Patologia Clínica, Faculdade de Ciências Médicas, Universidade Estadual de Campinas, 13083-970, Campinas, SP, Brazil

    Laura O. Saad, Sandra R. Mirandola, Evelise N. Maciel & Roger F. Castilho

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  1. Laura O. Saad
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  2. Sandra R. Mirandola
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  3. Evelise N. Maciel
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  4. Roger F. Castilho
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Correspondence to Roger F. Castilho.

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S. R. Mirandola and E. N. Maciel contributed equally to this work.

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Saad, L.O., Mirandola, S.R., Maciel, E.N. et al. Lactate Dehydrogenase Activity is Inhibited by Methylmalonate in vitro . Neurochem Res 31, 541–548 (2006). https://doi.org/10.1007/s11064-006-9054-6

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  • DOI: https://doi.org/10.1007/s11064-006-9054-6

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