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Top-down elasticity analysis and its application to energy metabolism in isolated mitochondria and intact cells

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Abstract

This paper reviews top-down elasticity analysis, which is a subset of metabolic control analysis. Top-down elasticity analysis provides a systematic yet simple experimental method to identify all the primary sites of action of an effector in complex systems and to distinguish them from all the secondary, indirect, sites of action. In the top-down approach, the complex system (for example, a mitochondrion, cell, organ or organism) is first conceptually divided into a small number of blocks of reactions interconnected by one or more metabolic intermediates. By changing the concentration of one intermediate when all others are held constant and measuring the fluxes through each block of reactions, the overall kinetic response of each block to each intermediate can be established. The concentrations of intermediates can be changed by adding new branches to the system or by manipulating the activities of blocks of reactions whose kinetics are not under investigation. To determine how much an effector alters the overall kinetics of a block of reactions, the overall kinetic response of the block to the intermediate is remeasured in the presence of the effector. Blocks that contain significant primary sites of action will display altered kinetics; blocks that change rate only because of secondary alterations in the concentrations of other metabolites will not. If desired, this elasticity analysis can be repeated with the primary target blocks subdivided into simpler blocks so that the primary sites of action can be defined with more and more precision until, with sufficient subdivision, they are mapped onto individual kinetic steps. Top-down elasticity analysis has been used to identify the targets of effectors of oxygen consumption in mitochondria, hepatocytes and thymocytes. Effectors include poisons such as cadmium and hormones such as tri-iodothyronine. However, the method is more general than this; in principle it can be applied to any metabolic or other steady-state system.

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References

  1. Hafner RP, Nobes CD, McGown AD, Brand MD: An altered relationship between proton motive force and respiration rate in liver mitochondria from hypothyroid rats shows that hypothyroidism decreases the proton leak (or slip). EBEC Short Rep 5: 128, 1988

    Google Scholar 

  2. Hafner RP, Nobes CD, MeGown AD, Brand MD: Altered relationship between proton motive force and respiration rate in non-phosphorylating liver mitochondria isolated from rats of different thyroid hormone status. Eur J Biochem 178: 511–518, 1988

    Google Scholar 

  3. Hafner RP, Brown GC, Brand MD: Thyroid-hormone control of state-3 respiration in isolated rat liver mitochondria. Biochem J 265: 731–734, 1990

    Google Scholar 

  4. Brand MD: The proton leak across the mitochondrial inner membrane. Biochim Biophys Acta 1018: 128–133, 1990

    Google Scholar 

  5. Nobes CD, Brown GC, Olive PN, Brand MD: Non-ohmic proton conductance of the mitochondrial inner membrane in hepatocytes. J Biol Chem 265: 12903–12909, 1990

    Google Scholar 

  6. Nobes CD, Hay WW, Brand MD: The mechanism of stimulation of respiration by fatty acid in isolated hepatocytes. J Biol Chem 265: 12910–12915, 1990

    Google Scholar 

  7. Brand MD, D'Allessandri L, Reis HMGPV, Hafner RP: Stimulation of the electron transport chain in mitochondria isolated from rats treated with mannoheptulose or glucagon. Arch Biochem Biophys 283: 278–284, 1990

    Google Scholar 

  8. Brand MD: Control of oxidative phosphorylation in liver mitochondria and cells: Top-down control analysis and top-down elasticity analysis. In: PW Hochachka, PL Lutz, T Sick, M Rosenthal, G van den Thillart (eds). Surviving Hypoxia: Mechanisms of Control and Adaptation. CRC Press, Boca Raton, 1993, pp 295–309

    Google Scholar 

  9. Harper M-E, Brand MD: Use of top-down elasticity analysis to identify sites of thyroid hormone-induced thermogenesis. Proc Soc Exp Biol Med 208: 228–237, 1995

    Google Scholar 

  10. Brand MD, Kesseler A: Control analysis of energy metabolism in mitochondria. Biochem Soc Trans 23: 371–376, 1995

    Google Scholar 

  11. Brand MD: Top down metabolic control analysis. J Theor Biol 182: 351–360, 1996

    Google Scholar 

  12. Brand MD: Regulation analysis of energy metabolism. J Exp Biol 200: 193–202, 1997

    Google Scholar 

  13. Kesseler A, Brand MD: Localisation of the sites of action of cadmium on oxidative phosphorylation in potato tuber mitochondria using top-down elasticity analysis. Eur J Biochem 225: 897–906, 1994

    Google Scholar 

  14. Ainscow EK, Brand MD: Top-down control analysis of systems with more than one common intermediate. Eur J Biochem 231: 579–586, 1995

    Google Scholar 

  15. Brown GC, Hafner RP, Brand MD: A ‘top-down’ approach to the determination of control coefficients in metabolic control theory. Eur J Biochem 188: 321–325, 1990

    Google Scholar 

  16. Brown GC: Phenomenological kinetics and the top-down approach to metabolic control analysis. In: S Schuster, M Rigoulet, R Ouhabi, J.-P. Mazat (eds). Modern Trends in Biothermokineties. Plenum Press, New York, 1994, pp 225–228

    Google Scholar 

  17. Kesseler A, Brand MD: Effects of cadmium on the control and internal regulation of oxidative phosphorylation in potato tuber mitochondria. Eur J Biochem 225: 907–922, 1994

    Google Scholar 

  18. Kholodenko BN, Brown GC: Paradoxical control properties of enzymes within pathways: Can activation cause an enzyme to have increased control? Biochem J 314: 753–760, 1996

    Google Scholar 

  19. Harper M-E, Brand MD: The quantitative contributions of mitochondrial proton leak and ATP turnover reactions to the changed respiration rates of hepatocytes from rats of different thyroid status. J Biol Chem 268: 14850–14860, 1993

    Google Scholar 

  20. Harper M-E, Ballantyne JS, Leach M, Brand MD: Effects of thyroid hormones on oxidative phosphorylation. Biochem Soc Trans 21: 785–792, 1993

    Google Scholar 

  21. Harper M-E, Brand MD: Mechanisms of the effects of hypothyroidism and hyperthyroidism in rats on respiration rate in isolated hepatocytes. In: S Schuster, M Rigoulet, R Ouhabi, J-P Mazat (eds). Modern Trends in Biothermokinetics. Plenum Press, New York, 1994, pp 351–356

    Google Scholar 

  22. Harper M-E, Brand MD: Hyperthyroidism stimulates mitochondrial proton leak and ATP turnover in rat hepatocytes but does not change the overall kinetics of substrate oxidation reactions. Can J Physiol Pharmacol 72: 899–908, 1994

    Google Scholar 

  23. Ainscow EK, Brand MD: Top-down control analysis of oxidative phosphorylation around ΔΨ and ATP in isolated rat hepatocytes. In: HV Westerhoff, JL Snoep, JE Wijker, FE Sluse, BN Kholodenko (eds). Biothermokinetics of the Living Cell. BioThermoKinetics Press, Amsterdam, 1996, pp 75–79

    Google Scholar 

  24. Nicholls DG: The influence of respiration and ATP hydrolysis on the proton-electrochemical gradient across the inner membrane of rat-liver mitochondria as determined by ion distribution.. Eur J Biochem 50: 305–315, 1974

    Google Scholar 

  25. Nicholls DG: The effective proton conductance of the inner membrane of mitochondria from brown adipose tissue. Dependency on proton electrochemical gradient. Eur J Biochem 77: 349–356, 1977

    Google Scholar 

  26. Westerhoff HV, van Dam K: Thermodynamics and Control of Biological Free Energy Transduction. Elsevier, Amsterdam, 1987

    Google Scholar 

  27. Brand MD, Steverding D, Kadenbach B, Stevenson PM, Hafner RP: The mechanism of the increase in mitochondrial proton permeability induced by thyroid hormones. Eur J Biochem 206: 775–781, 1992

    Google Scholar 

  28. Brown GC, Lakin-Thomas PL, Brand MD: Control of respiration and oxidative phosphorylation in isolated rat liver cells. Eur J Biochem 192: 355–362, 1990

    Google Scholar 

  29. Buttgereit F, Grant A, Mtiller M, Brand MD: The effects of methylprednisolone on oxidative phosphorylation in Concanavalin-Astimulated thymocytes. Top down elasticity analysis and control analysis. Eur J Biochem 223: 513–519, 1994

    Google Scholar 

  30. Brown GC, Brand MD: On the nature of the mitochondrial proton leak. Biochim Biophys Acta 1059: 55–62, 1991

    Google Scholar 

  31. Brand MD, Couture P, Else PL, Withers KW, Hulbert AJ: Evolution of energy metabolism: Proton permeability of the inner membrane of liver mitochondria is greater in a mammal than in a reptile. Biochem J 275: 81–86, 1991

    Google Scholar 

  32. Porter RK, Brand MD: The contribution of mitochondrial proton leak to basal metabolic rate: A comparison of the proton leak kinetics in mammals of different body mass. EBEC Reports 7: 63, 1992

    Google Scholar 

  33. Porter RK, Brand MD: Body mass dependence of H+ leak in mitochondria and its relevance to metabolic rate. Nature 362: 628–630, 1993

    Google Scholar 

  34. Porter RK, Brand MD: Causes of the differences in respiration rate of hepatocytes from mammals of different body mass. Am J Physiol 269: R1213–R1224, 1995

    Google Scholar 

  35. Rolfe DFS, Hulbert AJ, Brand MD: Characteristics of mitochondrial proton leak and control of oxidative phosphorylation in the major oxygen consuming tissues of the rat. Biochim Biophys Acta 1188: 405–416, 1994

    Google Scholar 

  36. Kesseler A, Diolez P, Brinkman K, Brand MD: Characterisation of the control of respiration in potato tuber mitochondria using the top-down approach of metabolic control analysis. Eur J Biochem 210: 775–784, 1992

    Google Scholar 

  37. Brand MD, Chien L-F, Rolfe DFS: Control of oxidative phosphorylation in liver mitochondria and hepatocytes. Biochem Soc Trans 21: 757–762, 1993

    Google Scholar 

  38. Fusi F, Sgaragli G, Murphy MP: Interaction of butylated hydroxyanisole with mitochondrial oxidative phosphorylation. Biochem Pharm 43: 1203–1208, 1992

    Google Scholar 

  39. Makins RA, Drynan LF, Zammit VA, Quant PA: Top-down regulation analyses of palmitoyl-CoA oxidation and ketogenesis in isolated rat liver mitochondria. Biochem Soc Trans 23: 288S, 1995

    Google Scholar 

  40. Mildaziene V, Marcinkeviciute A, Baniene R, Mazat J-P: Supraphysiological concentration of calcium inhibits phosphorylation, respiratory subsystem and uncouples rat heart mitochondria. In: HV Westerhoff, JL Snoep, JE Wijker, FE Sluse, BN Kholodenko (eds). Biothermokinetics of the Living Cell. BioThermoKinetics Press, Amsterdam, 1996, pp 168–173

    Google Scholar 

  41. Kesseler A, Brand MD: The mechanism of the stimulation of state 4 respiration by cadmium in potato tuber (Solanum tuberosum) mitochondria. Plant Physiol Biochem 33: 519–528, 1995

    Google Scholar 

  42. Borutaite V, Mildaziene V, Brown GC, Brand MD: Control and kinetic analysis of ischemia-damaged heart mitochondria: Which parts of the oxidative phosphorylation system are affected by ischemia? Biochim Biophys Acta 1272: 154–158, 1995

    Google Scholar 

  43. Borutaite V, Morkuniene R, Budriunaite A, Brown GC: Phenomenological kinetic analysis of ischemic damage to heart mitochondria: Role of Ca2+, fatty acids and cytochrome c. In: HV Westerhoff, JL Snoep, JE Wijker, FE Sluse, BN Kholodenko (eds). Biothermokinetics of the Living Cell. BioThermoKinetics Press, Amsterdam, 1996, pp 89–93

    Google Scholar 

  44. Lionetti L, Iossa S, Brand MD, Liverini G: Relationship between membrane potential and respiration rate in isolated liver mitochondria from rats fed an energy dense diet. Mol Cell Biochem 158: 133–138, 1996

    Google Scholar 

  45. Lionetti L, Iossa S, Brand MD, Liverini G: The mechanism of stimulation of respiration in isolated hepatocytes from rats fed an energy dense diet. J Nutr Biochem 7: 571–576, 1996

    Google Scholar 

  46. Dufour S, Rousse N, Canioni P, Diolez P: Top-down control analysis of temperature effect on oxidative phosphorylation. Biochem J 314: 743–751, 1996

    Google Scholar 

  47. Dufour S, Rousse N, Canioni P, Diolez P: Study of a complex effector of oxidative phosphorylation using top down elasticity analysis and control analysis. In: HV Westerhoff, JL Snoep, JE Wijker, FE Sluse, BN Kholodenko (eds). Biothermokinetics of the Living Cell. BioThermoKinetics Press, Amsterdam, 1996, pp 26–30

    Google Scholar 

  48. Oliver SG: From DNA sequence to biological function. Nature 379: 697–600, 1996

    Google Scholar 

  49. Dujon B: The yeast genome project: What did we learn? Trends Genet 12: 263–270, 1996

    Google Scholar 

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  1. Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK

    Martin D. Brand

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Brand, M.D. Top-down elasticity analysis and its application to energy metabolism in isolated mitochondria and intact cells. Mol Cell Biochem 184, 13–20 (1998). https://doi.org/10.1023/A:1006893619101

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