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Production and export of metabolites from liver and heart mitochondria and anaplerosis

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Abstract

A method to study the export of citric acid cycle intermediates from rat liver mitochondria supplied with various individual substrates or combinations of substrates was designed to focus on the role of mitochondria in anaplerosis and cataplerosis. Under most conditions malate, citrate, and aspartate were exported in far higher amounts than isocitrate and α-ketoglutarate. In the presence of pyruvate alone or pyruvate in combination with most other substrates, citrate export equaled or was only slightly less than malate export. This contrasts with pancreatic islet mitochondria where citrate export is unaffected by many substrates. Malate and succinate potentiated pyruvate-induced citrate export and succinate caused massive malate export from liver mitochondria. Heart mitochondria, which possess very little or no pyruvate carboxylase, unlike liver and pancreatic islet mitochondria, did not produce malate from pyruvate. Heart mitochondria produced malate, but not citrate, from succinate. The results indicate that liver mitochondria export a larger number of metabolites from a wider range of substrates than do islet or heart mitochondria. This may reflect the multiple roles of the liver in body metabolism versus the specialized roles of the islet cell and heart.

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References

  1. MacDonald MJ: Feasibility of a mitochondrial pyruvate malate shuttle in pancreatic islets. J Biol Chem 270: 20051–20058, 1995

    Google Scholar 

  2. Owen OE, Kalhan SC, Hanson RW: The key role of anaplerosis and cataplerosis for citric acid cycle function. J Biol Chem 277: 30409–30412, 2002

    Google Scholar 

  3. Walter P, Paetkau V, Lardy HA: Paths of carbon in gluconeogenesis and lipogenesis. 3. The role and regulation of mitochondrial processes involved in supplying precursors of phosphoenolpyruvate. J Biol Chem 241: 2523–2532, 1966

    Google Scholar 

  4. Bobyleva V, Kneer N, Bellei M, Battelli D, Lardy HA: Concerning the mechanism of increased thermogenesis in rats treated with dehydroepiandrosterone. J Bioenergetics Biomembr 25: 313–332, 1993

    Google Scholar 

  5. Lardy HA, Paetkau V, Walter P: Paths of carbon in gluconeogenesis and lipogenesis: the role of mitochondria in supplying precursors of phosphoenolpyruvate. Proc Natl Acad Sci USA 53: 1410–1415, 1965

    Google Scholar 

  6. Wiese TJ, Wuensch SA, Ray PD: Synthesis of citrate from phosphoenolpyruvate and acetylcarnitine by mitochondria from rabbit, pigeon and rat liver: Implications for lipogenesis. Comp Biochem Physiol 114B: 417–422, 1996

    Google Scholar 

  7. MacDonald MJ, Chang C-M: Do pancreatic islets contain significant amounts of phosphoenol-pyruvate carboxykinase or ferroactivator activity? Diabetes 34: 246–250, 1985

    Google Scholar 

  8. MacDonald, MJ, McKenzie DI, Walker TM, Kaysen JH: Lack of glyconeogenesis in pancreatic islets: Expression of gluconeogenic enzyme genes in islets. Hormone Metab Res 24: 158–160, 1992

    Google Scholar 

  9. MacDonald MJ: Metabolism of the insulin secretagogue methyl succinate by pancreatic islets. Arch Biochem Biophys 300: 201–205, 1993

    Google Scholar 

  10. MacDonald MJ: Estimates of glycolysis, pyruvate (De)carboxylation, pentose phosphate pathway and methyl succinate metabolism in incapacitated pancreatic islets. Arch Biochem Biophys 305: 205–214, 1993

    Google Scholar 

  11. Khan A, Ling ZC, Landau BR: Quantifying the carboxylation of pyruvate in pancreatic islets. J Biol Chem 271: 2539–2542, 1996

    Google Scholar 

  12. Lu D, Mulder H, Zhao P, Burgess SC, Jensen MV, Kamzolova S, Newgard CB, Sherry AD: 13C NMR isotopomer analysis reveals a connection between pyruvate cycling and glucose-stimulated insulin secretion (GSIS). Proc Natl Acad Sci 99: 2708–2713, 2002

    Google Scholar 

  13. MacDonald MJ: The export of metabolites from mitochondria and anaplerosis in insulin secretion. Biochim Biophys Acta 1619: 77–88, 2003

    Google Scholar 

  14. Davis EJ, Spydevoid O, Bremer J: Pyruvate carboxylase and propionyl-CoA carboxylase as anaplerotic enzymes in skeletal muscle mitochondria. Eur J Biochem 110: 255–262, 1980

    Google Scholar 

  15. Henslee JG, Srere PA: Resolution of rat mitochondrial matrix proteins by two-dimensional polyacrylamide gel electrophoresis. J Biol Chem 254: 5488–5497, 1979

    Google Scholar 

  16. MacDonald MJ: Evidence for the malate-asparate shuttle in pancreatic islets. Arch Biochem Biophys 213: 643–649, 1982

    Google Scholar 

  17. Johnson D, Lardy HA: Isolation of liver or kidney mitochondria. Meth Enzymol 10: 94–96, 1967

    Google Scholar 

  18. Passonneau JV, Lowry OH: In: Enzymatic Analysis, A Practical Guide. Humana Press, Totowa, NJ, 1993

    Google Scholar 

  19. Bergmeyer HU, Bergmeyer J, Grassl M: In: Methods of Enzymatic Analysis, 3rd edn, vol. 7; Metabolites 2: VCH, Verlagsgesellshaft, Weinheim, FRG, 1984

    Google Scholar 

  20. MacDonald MJ, Neufelt N, Park B, Berger M, Ruderman NB: Alanine metabolism and glucoenogenesis in the rat. Am J Physiol 231: 619–626, 1976

    Google Scholar 

  21. MacDonald MJ, Huang MT, Lardy HA: Hyperglycemic activity and metabolic effects of 3-aminopicolinic acid. Biochem J 176: 495–504, 1978

    Google Scholar 

  22. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ: Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275, 1951

    Google Scholar 

  23. MacDonald MJ: Differences between mouse and rat pancreatic islets: Succinate responsiveness, malic enzyme and anaplerosis. Am J Physiol Endo Metab 283: E302–E310, 2002

    Google Scholar 

  24. Atkinson DE: In: J.M. Lowenstein (ed). Citric Acid Cycle: Control and Compartmentation. Marcel Dekker, New York, 1969, pp 137–162

    Google Scholar 

  25. MacDonald MJ, Al-Masri H, Jumelle-Laclau M, Cruz M: Oscillations in activities of enzymes in pancreatic islet subcellular fractions induced by physiological concentrations of effectors. Diabetes 46: 1996–2001, 1997

    Google Scholar 

  26. Fahien LA, MacDonald MJ, Kmiotek EH, Mertz RJ, Fahien CM: Regulation of insulin release by factors that also modify glutamate dehydrogenase. J Biol Chem 263: 13610–13614, 1988

    Google Scholar 

  27. Gylfe E: Comparison of the effects of leucines, non-metabolizable leucine analogues and other insulin secretagogues on the activity of glutamate dehydrogenase. Acta Diabetol Lat 13: 20–24, 1976

    Google Scholar 

  28. Malaisse-Lagae F, Sener A, Garcia-Morales P, Valverde I, Malaisse WJ: The stimulus-secretion coupling of amino acid-induced insulin release. Influence of a nonmetabolized analog of leucine on the metabolism of glutamine in pancreatic islets. J Biol Chem 257: 3754–3758, 1982

    Google Scholar 

  29. Lenzen S, Schmidt W, Panten U: Transamination of neutral amino acids and 2-keto acids in pancreatic B-cell mitochondria. J Biol Chem 260: 12629–12634, 1985

    Google Scholar 

  30. MacDonald MJ, Fahien LA: Glutamate is not a messenger in insulin secretion. J Biol Chem 275: 34025–34027, 2000

    Google Scholar 

  31. Fahien LA, Teller JK: Glutamate-malate metabolism in liver mitochondria. A model constructed on the basis of mitochondrial levels of enzymes, specificity, dissociation constants, and stoichiometry of hetero-enzyme complexes. J Biol Chem 267: 10411–10422, 1992

    Google Scholar 

  32. Williamson DH, Lund P, Krebs HA: The redox state of free nicotinamide-adenine dinucleotide in the cytoplasm and mitochondria of rat liver. Biochem J 103: 514–527, 1967

    Google Scholar 

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Authors and Affiliations

  1. University of Wisconsin Childrens Diabetes Center, Madison, WI, 53706, USA

    Michael J. MacDonald

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  1. Michael J. MacDonald
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MacDonald, M.J. Production and export of metabolites from liver and heart mitochondria and anaplerosis. Mol Cell Biochem 258, 201–210 (2004). https://doi.org/10.1023/B:MCBI.0000012856.31929.91

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  • DOI: https://doi.org/10.1023/B:MCBI.0000012856.31929.91

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