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Intramitochondrial localization of alanine aminotransferase in rat-liver mitochondria: comparison with glutaminase and aspartate aminotransferase

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Summary

The removal of the outer mitochondrial membrane and hence of constituents of the intermembrane space in rat-liver mitochondria using digitonin showed that phosphate-dependent glutaminase, alanine and aspartate aminotransferase were localized in the mitoplasts. Further fractionation of mitoplasts following their sonication resulted in 90% of glutaminase, 98% of alanine aminotransferase and 48% of aspartate aminotransferase being recovered in the soluble fraction while the remainder of each enzyme was recovered in the sonicated vesicles fraction. These results indicated that glutaminase and alanine aminotransferase were soluble matrix enzymes, the little of each enzyme recovered in the sonicated vesicles fraction being probably due to entrapment in the vesicles. Aspartate aminotransferase had dual localization, in the inner membrane and matrix with the high specific activity in sonicated vesicles confirming its association with the membrane. Activation experiments suggested that the membrane-bound enzyme was localized on the inner side of the inner mitochondrial membrane.

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References

  • Azzi A, Chappell JB, Robinson BH (1967) Penetration of the mitochondrial membrane by glutamate and aspartate. Biochim Biophys Res Commun 29: 148–152

    Google Scholar 

  • Bergmeyer HU (1974) Myokinase, adenylate kinase from rabbit muscle. In: Methods of enzymatic analysis, vol 1, 2nd edn. Academic Press, New York London, pp 486–487

    Google Scholar 

  • Boyd JW (1961) The intracellular distribution, latency and electrophoretic mobility of L-glutamate-oxaloacetate transaminase from rat liver. Biochem J 81: 434–441

    Google Scholar 

  • Brdiczka D, Pette D, Brunner G, Miller F (1968) Compartmental distribution of enzymes in rat liver mitochondria. Eur J Biochem 5: 294–304

    Google Scholar 

  • Chappell JB, Hansford RG (1972) Preparation of mitochondria from animal tissues and yeasts. In: Bernie GD (ed) Subcellular components, preparation and fractionation. In: Butterworth, London, pp 77–91

    Google Scholar 

  • Comte J, Gautheron DC (1978) The markers of pig heart mitochondrial sub-fractions. II. On the association of malate dehydrogenase with inner membrane. Biochimie 60: 1299–1305

    Google Scholar 

  • Curthoys NP, Weiss RF (1974) Regulation of renal ammoniagenesis. Subcelluar localisation of rat kidney glutaminase isoenzymes. J Biol Chem 249: 3261–3266

    Google Scholar 

  • DeRosa G, Swick RW (1975) Metabolic implications of the distribution of the alanine aminotransferase isoenzymes. J Biol Chem 250: 7961–7967

    Google Scholar 

  • Duszynski J, Mueller G, LaNoue K (1978) Microcompartmentation of aspartate in rat liver mitochondria. J Biol Chem 253: 6149–6157

    Google Scholar 

  • Elduque A, Casadó F, Cortés A, Bozal J (1982) Intramitochondrial location of the molecular forms of chicken liver mitochondrial malate dehydrogenase. Int J Biochem 14: 221–229

    Google Scholar 

  • Evered DF, Masola B (1984) The oxidation of glutamine and glutamate in relation to anion transport in enterocyte mitochondria. Biochem J 218: 449–458

    Google Scholar 

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

    Google Scholar 

  • Fleisher GA, Potter CS, Wakim KG (1960) Separation of two glutamic oxaloacetic transaminases by paper electrophoresis. Proc Soc Exptl Biol Med 103: 229–231

    Google Scholar 

  • Gil M, Cascante M, Cortés A, Bozal J (1987) Intramitochondrial location and some characteristics of chicken liver aspartate aminotransferase. Int J Biochem 19: 355–363

    Google Scholar 

  • Gornall AG, Bardawill CS, David MM (1949) Determination of serum proteins by means of the biuret reaction. J Biol Chem 177: 751–766

    Google Scholar 

  • Gotterer GS (1967) Rat liver D-β-hydroxybutyrate dehydrogenase. I. Partial purification and general properties. Biochemistry 6: 2139–2152

    Google Scholar 

  • Groen AK, Sips HJ, Vervoorn RC, Tager JM (1982) Intracellular compartmentation and control of alanine metabolism in rat liver parenchymal cells. Eur J Biochem 122: 87–93

    Google Scholar 

  • Harmon HJ, Hall JD, Crane FL (1974) Structure of mitochondrial cristae membranes. Biochim Biophys Acta 344: 119–155

    Google Scholar 

  • Huang CH, Keyhani E, Lee CP (1973) Fractionation by sucrose density gradient centrifugation of membrane fragments derived by sonic disruption of beef heart mitochondria. Biochim Biophys Acta 305: 455–473

    Google Scholar 

  • Kovačević Z (1971) The pathway of glutamine and glutamate oxidation in isolated mitochondria from mammalian cells. Biochem J 125: 757–763

    Google Scholar 

  • Kvamme E, Torgner IA, Roberg B (1991) Evidence indicating that pig renal phosphate-activated glutaminase has a functionally predominant external localisation in the inner mitochondrial membrane. J Biol Chem 266: 13185–13192

    Google Scholar 

  • Landriscina C, Papa S, Corattelli P, Mazzarella L, Quangliariello E (1970) Enzyme activities of the matrix and inner membrane of pigeon-liver mitochondria. Biochim Biophys Acta 205: 136–147

    Google Scholar 

  • LaNoue KF, Tischler ME (1974) Electrogenic characteristics of the mitochondrial glutamate-aspartate antiporter. J Biol Chem 248: 7522–7528

    Google Scholar 

  • Lenartowicz E, Wojtczak AB (1988) Significance of the alanine aminotransferase reaction in the formation ofα-ketoglutarate in rat liver mitochondria. Arch Biochem Biophys 260: 309–319

    Google Scholar 

  • Maïrouch H, Godinot C (1977) Localisation of protein(s) involved in oligomycin — induced inhibition of mitochondrial adenosine triphosphatase near the outer surface of the inner membrane. Proc Natl Acad Sci USA 74: 4185–4189

    Google Scholar 

  • Maisterrena B, Comte J, Gautheron DC (1974) Purification of pig heart mitochondrial membranes enzymatic and morphological characterisation as compared to microsomes. Biochim Biophys Acta 367: 115–126

    Google Scholar 

  • Marco R, Sebastián J, Sols A (1969) Location of the enzymes of the oxaloacetate metabolic cross-roads in rat liver mitochondria. Biochem Biophys Res Commun 34: 725–730

    Google Scholar 

  • Masola B, Peters TJ, Evered DF (1985) Transamination pathways influencing L-glutamine and L-glutamate oxidation by rat enterocyte mitochondria and the subcelluar localization of L-alanine aminotransferase and L-aspartate aminotransferase. Biochem Biophys Acta 843: 137–143

    Google Scholar 

  • McGivan JD, Lacey JH, Joseph SK (1980) Localisation and some properties of phosphate-dependent glutaminase in disrupted mitochondria. Biochem J 192: 537–542

    Google Scholar 

  • McIntyre JO, Bock H-GO, Fleischer S (1978) The orientation of D-β-hydroxybutyrate dehydrogenase in the mitochondrial inner membrane. Biochim Biophys Acta 513: 255–267

    Google Scholar 

  • Pinkus LM, Windmueller HG (1977) Phosphate-dependent glutaminase of small intestine: localisation and role in intestinal glutamine metabolism. Arch Biochem Biophys 182: 507–517

    Google Scholar 

  • Pour-Rahimi F, Nemat-Gorgani M (1987) Reversible association of ox liver glutamate dehydrogenase with the inner mitochondrial membrane. Int J Biochem 19: 53–61

    Google Scholar 

  • Sattocasa GL, Kuylenstierna B, Ernster L, Bergstrand A (1967) An electron-transport associated with the outer membrane of liver mitochondria. J Cell Biol 32: 415–438

    Google Scholar 

  • Schmidt E (1974) Glutamate dehydrogenase U.V. assay. In: Bergmeyer HU (ed) Methods of enzymatic analysis, vol. 2, 2nd edn. Academic Press, New York, pp 650–656

    Google Scholar 

  • Schnaitaman C, Erwin VG, Greenawalt JW (1967) The submitochondrial localisation of monoamine oxidase. An enzymatic marker for the outer membrane of rat liver mitochondria. J Cell Biol 32: 719–735

    Google Scholar 

  • Schnaitman C, Greenawalt JW (1968) Enzymatic properties of the inner and outer membranes of rat liver mitochondrial. J Cell Biol 38: 158–175

    Google Scholar 

  • Shapiro RA, Haser WG, Curthoys NP (1985) The orientation of phosphate-dependent glutaminase on the inner membrane of rat renal mitochondria. Arch Biochem Biophys 243: 1–7

    Google Scholar 

  • Swick RW, Barnstein PL, Stange JL (1965) The metabolism of mitochondrial proteins. I. Distribution and characterisation of the isoenzymes of alanine aminotransferase in rat liver. J Biol Chem 240: 3334–3340

    Google Scholar 

  • Tabor CW, Tabor H, Rosenthal SM (1954) Purification of amine oxidase from beef plasma. J Biol Chem 208: 645–651

    Google Scholar 

  • Teller JK, Fahien LA, Valdivia E (1990) Interactions among mitochondrial aspartate aminotransferase, malate dehydrogenase, and the inner mitochondrial membrane from heart, hepatoma, and liver. J Biol Chem 265: 19486–19494

    Google Scholar 

  • Volman-Mitchell H, Parson DS (1974) Distribution and activities of dicarboxylic amino acid transaminases in gastrointestinal mucosa of rat, mouse, hamster, guinea pig, chicken and pigeon. Biochim Biophys Acta 334: 316–327

    Google Scholar 

  • Waksman A, Rendon A (1974) Intramitochondrial intermembranal large amplitude protein movements. Biochimie 56: 907–924

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, University of Zimbabwe, P.O. Box MP 167, Mount Pleasant, Harare, Zimbabwe

    B. Masola

  2. Department of Biological Chemistry, Hahnemann University, Philadelphia, Pennsylvania, USA

    T. M. Devlin

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  1. B. Masola
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  2. T. M. Devlin
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Masola, B., Devlin, T.M. Intramitochondrial localization of alanine aminotransferase in rat-liver mitochondria: comparison with glutaminase and aspartate aminotransferase. Amino Acids 9, 363–374 (1995). https://doi.org/10.1007/BF00807273

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  • DOI: https://doi.org/10.1007/BF00807273

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