• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of biochemjBJ Latest papers and much more!
Biochem J. May 15, 1988; 252(1): 181–189.
PMCID: PMC1149122

Regulation of NAD+-linked isocitrate dehydrogenase and 2-oxoglutarate dehydrogenase by Ca2+ ions within toluene-permeabilized rat heart mitochondria. Interactions with regulation by adenine nucleotides and NADH/NAD+ ratios.

Abstract

1. Toluene-permeabilized rat heart mitochondria have been used to study the regulation of NAD+-linked isocitrate dehydrogenase and 2-oxoglutarate dehydrogenase by Ca2+, adenine and nicotinamide nucleotides, and to compare the properties of the enzymes in situ, with those in mitochondrial extracts. 2. Although K0.5 values (concn. giving half-maximal effect) for Ca2+ of 2-oxoglutarate dehydrogenase were around 1 microM under all conditions, corresponding values for NAD+-linked isocitrate dehydrogenase were in the range 5-43 microM. 3. For both enzymes, K0.5 values for Ca2+ observed in the presence of ATP were 3-10-fold higher than those in the presence of ADP, with values increasing over the ADP/ATP range 0.0-1.0. 4. 2-Oxoglutarate dehydrogenase was less sensitive to inhibition by NADH when assayed in permeabilized mitochondria than in mitochondrial extracts. Similarly, the Km of NAD+-linked isocitrate dehydrogenase for threo-Ds-isocitrate was lower in permeabilized mitochondria than in extracts under all the conditions investigated. 5. It is concluded that in the intact heart Ca2+ activation of NAD+-linked isocitrate dehydrogenase may not necessarily occur in parallel with that of the other mitochondrial Ca2+-sensitive enzymes, 2-oxoglutarate dehydrogenase and the pyruvate dehydrogenase system.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Akerboom TP, Bookelman H, Zuurendonk PF, van der Meer R, Tager JM. Intramitochondrial and extramitochondrial concentrations of adenine nucleotides and inorganic phosphate in isolated hepatocytes from fasted rats. Eur J Biochem. 1978 Mar 15;84(2):413–420. [PubMed]
  • Balaban RS, Kantor HL, Katz LA, Briggs RW. Relation between work and phosphate metabolite in the in vivo paced mammalian heart. Science. 1986 May 30;232(4754):1121–1123. [PubMed]
  • Belsham GJ, Denton RM, Tanner MJ. Use of a novel rapid preparation of fat-cell plasma membranes employing Percoll to investigate the effects of insulin and adrenaline on membrane protein phosphorylation within intact fat-cells. Biochem J. 1980 Nov 15;192(2):457–467. [PMC free article] [PubMed]
  • Brand MD, Murphy MP. Control of electron flux through the respiratory chain in mitochondria and cells. Biol Rev Camb Philos Soc. 1987 May;62(2):141–193. [PubMed]
  • Corkey BE, Duszynski J, Rich TL, Matschinsky B, Williamson JR. Regulation of free and bound magnesium in rat hepatocytes and isolated mitochondria. J Biol Chem. 1986 Feb 25;261(6):2567–2574. [PubMed]
  • Crompton M, Kessar P, Al-Nasser I. The alpha-adrenergic-mediated activation of the cardiac mitochondrial Ca2+ uniporter and its role in the control of intramitochondrial Ca2+ in vivo. Biochem J. 1983 Nov 15;216(2):333–342. [PMC free article] [PubMed]
  • Denton RM, McCormack JG. On the role of the calcium transport cycle in heart and other mammalian mitochondria. FEBS Lett. 1980 Sep 22;119(1):1–8. [PubMed]
  • Denton RM, McCormack JG. Ca2+ transport by mammalian mitochondria and its role in hormone action. Am J Physiol. 1985 Dec;249(6 Pt 1):E543–E554. [PubMed]
  • Denton RM, Randle PJ, Martin BR. Stimulation by calcium ions of pyruvate dehydrogenase phosphate phosphatase. Biochem J. 1972 Jun;128(1):161–163. [PMC free article] [PubMed]
  • Denton RM, Richards DA, Chin JG. Calcium ions and the regulation of NAD+-linked isocitrate dehydrogenase from the mitochondria of rat heart and other tissues. Biochem J. 1978 Dec 15;176(3):899–906. [PMC free article] [PubMed]
  • Denton RM, McCormack JG, Edgell NJ. Role of calcium ions in the regulation of intramitochondrial metabolism. Effects of Na+, Mg2+ and ruthenium red on the Ca2+-stimulated oxidation of oxoglutarate and on pyruvate dehydrogenase activity in intact rat heart mitochondria. Biochem J. 1980 Jul 15;190(1):107–117. [PMC free article] [PubMed]
  • From AH, Petein MA, Michurski SP, Zimmer SD, Uğurbil K. 31P-NMR studies of respiratory regulation in the intact myocardium. FEBS Lett. 1986 Oct 6;206(2):257–261. [PubMed]
  • Gabriel JL, Plaut GW. Inhibition of bovine heart NAD-specific isocitrate dehydrogenase by reduced pyridine nucleotides: modulation of inhibition by ADP, NAD+, Ca2+, citrate, and isocitrate. Biochemistry. 1984 Jun 5;23(12):2773–2778. [PubMed]
  • Gabriel JL, Milner R, Plaut GW. Inhibition and activation of bovine heart NAD-specific isocitrate dehydrogenase by ATP. Arch Biochem Biophys. 1985 Jul;240(1):128–134. [PubMed]
  • Gabriel JL, Zervos PR, Plaut GW. Activity of purified NAD-specific isocitrate dehydrogenase at modulator and substrate concentrations approximating conditions in mitochondria. Metabolism. 1986 Jul;35(7):661–667. [PubMed]
  • Halestrap AP. The regulation of the oxidation of fatty acids and other substrates in rat heart mitochondria by changes in the matrix volume induced by osmotic strength, valinomycin and Ca2+. Biochem J. 1987 May 15;244(1):159–164. [PMC free article] [PubMed]
  • Hansford RG. Relation between mitochondrial calcium transport and control of energy metabolism. Rev Physiol Biochem Pharmacol. 1985;102:1–72. [PubMed]
  • Hansford RG, Castro F. Intramitochondrial and extramitochondrial free calcium ion concentrations of suspensions of heart mitochondria with very low, plausibly physiological, contents of total calcium. J Bioenerg Biomembr. 1982 Dec;14(5-6):361–376. [PubMed]
  • Johnston JD, Brand MD. Stimulation of the respiration rate of rat liver mitochondria by sub-micromolar concentrations of extramitochondrial Ca2+. Biochem J. 1987 Jul 1;245(1):217–222. [PMC free article] [PubMed]
  • Jung DW, Brierley GP. Matrix magnesium and the permeability of heart mitochondria to potassium ion. J Biol Chem. 1986 May 15;261(14):6408–6415. [PubMed]
  • Katz LA, Koretsky AP, Balaban RS. Respiratory control in the glucose perfused heart. A 31P NMR and NADH fluorescence study. FEBS Lett. 1987 Sep 14;221(2):270–276. [PubMed]
  • Kendrick NC. Purification of arsenazo III, a Ca2+-sensitive dye. Anal Biochem. 1976 Dec;76(2):487–501. [PubMed]
  • Lawlis VB, Roche TE. Effect of micromolar Ca2+ on NADH inhibition of bovine kidney alpha-ketoglutarate dehydrogenase complex and possible role of Ca2+ in signal amplification. Mol Cell Biochem. 1980 Nov 20;32(3):147–152. [PubMed]
  • Lawlis VB, Roche TE. Inhibition of bovine kidney alpha-ketoglutarate dehydrogenase complex by reduced nicotinamide adenine dinucleotide in the presence or absence of calcium ion and effect of adenosine 5'-diphosphate on reduced nicotinamide adenine dinucleotide inhibition. Biochemistry. 1981 Apr 28;20(9):2519–2524. [PubMed]
  • Linn TC, Pelley JW, Pettit FH, Hucho F, Randall DD, Reed LJ. -Keto acid dehydrogenase complexes. XV. Purification and properties of the component enzymes of the pyruvate dehydrogenase complexes from bovine kidney and heart. Arch Biochem Biophys. 1972 Feb;148(2):327–342. [PubMed]
  • Marshall SE, McCormack JG, Denton RM. Role of Ca2+ ions in the regulation of intramitochondrial metabolism in rat epididymal adipose tissue. Evidence against a role for Ca2+ in the activation of pyruvate dehydrogenase by insulin. Biochem J. 1984 Feb 15;218(1):249–260. [PMC free article] [PubMed]
  • Matlib MA, Shannon WA, Jr, Srere PA. Measurement of matrix enzyme activity in isolated mitochondria made permeable with toluene. Arch Biochem Biophys. 1977 Jan 30;178(2):396–407. [PubMed]
  • McCormack JG. Characterization of the effects of Ca2+ on the intramitochondrial Ca2+-sensitive enzymes from rat liver and within intact rat liver mitochondria. Biochem J. 1985 Nov 1;231(3):581–595. [PMC free article] [PubMed]
  • McCormack JG, Denton RM. The effects of calcium ions and adenine nucleotides on the activity of pig heart 2-oxoglutarate dehydrogenase complex. Biochem J. 1979 Jun 15;180(3):533–544. [PMC free article] [PubMed]
  • McCormack JG, Denton RM. Role of calcium ions in the regulation of intramitochondrial metabolism. Properties of the Ca2+-sensitive dehydrogenases within intact uncoupled mitochondria from the white and brown adipose tissue of the rat. Biochem J. 1980 Jul 15;190(1):95–105. [PMC free article] [PubMed]
  • McCormack JG, Denton RM. A comparative study of the regulation of Ca2+ of the activities of the 2-oxoglutarate dehydrogenase complex and NAD+-isocitrate dehydrogenase from a variety of sources. Biochem J. 1981 May 15;196(2):619–624. [PMC free article] [PubMed]
  • Midgley PJ, Rutter GA, Thomas AP, Denton RM. Effects of Ca2+ and Mg2+ on the activity of pyruvate dehydrogenase phosphate phosphatase within toluene-permeabilized mitochondria. Biochem J. 1987 Jan 15;241(2):371–377. [PMC free article] [PubMed]
  • Miller DJ, Smith GL. EGTA purity and the buffering of calcium ions in physiological solutions. Am J Physiol. 1984 Jan;246(1 Pt 1):C160–C166. [PubMed]
  • Neely JR, Denton RM, England PJ, Randle PJ. The effects of increased heart work on the tricarboxylate cycle and its interactions with glycolysis in the perfused rat heart. Biochem J. 1972 Jun;128(1):147–159. [PMC free article] [PubMed]
  • Scharschmidt BF, Keeffe EB, Blankenship NM, Ockner RK. Validation of a recording spectrophotometric method for measurement of membrane-associated Mg- and NaK-ATPase activity. J Lab Clin Med. 1979 May;93(5):790–799. [PubMed]
  • Siess EA, Brocks DG, Lattke HK, Wieland OH. Effect of glucagon on metabolite compartmentation in isolated rat liver cells during gluconeogenesis from lactate. Biochem J. 1977 Aug 15;166(2):225–235. [PMC free article] [PubMed]
  • Smith CM, Bryla J, Williamson JR. Regulation of mitochondrial alpha-ketoglutarate metabolism by product inhibition at alpha-ketoglutarate dehydrogenase. J Biol Chem. 1974 Mar 10;249(5):1497–1505. [PubMed]
  • Soboll S, Bünger R. Compartmentation of adenine nucleotides in the isolated working guinea pig heart stimulated by noradrenaline. Hoppe Seylers Z Physiol Chem. 1981 Feb;362(2):125–132. [PubMed]
  • Soboll S, Akerboom TP, Schwenke WD, Haase R, Sies H. Mitochondrial and cytosolic ATP/ADP ratios in isolated hepatocytes. A comparison of the digitonin method and the non-aqueous fractionation procedure. Biochem J. 1980 Dec 15;192(3):951–954. [PMC free article] [PubMed]
  • Soboll S, Seitz HJ, Sies H, Ziegler B, Scholz R. Effect of long-chain fatty acyl-CoA on mitochondrial and cytosolic ATP/ADP ratios in the intact liver cell. Biochem J. 1984 Jun 1;220(2):371–376. [PMC free article] [PubMed]
  • Thomas AP, Denton RM. Use of toluene-permeabilized mitochondria to study the regulation of adipose tissue pyruvate dehydrogenase in situ. Further evidence that insulin acts through stimulation of pyruvate dehydrogenase phosphate phosphatase. Biochem J. 1986 Aug 15;238(1):93–101. [PMC free article] [PubMed]
  • Thomas AP, Diggle TA, Denton RM. Sensitivity of pyruvate dehydrogenase phosphate phosphatase to magnesium ions. Similar effects of spermine and insulin. Biochem J. 1986 Aug 15;238(1):83–91. [PMC free article] [PubMed]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

Formats:

Related citations in PubMed

Cited by other articles in PMC

See all...

Links

  • Compound
    Compound
    PubChem Compound links
  • MedGen
    MedGen
    Related information in MedGen
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...