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2007 | 54 | 4 | 797-803
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Response of Acanthamoeba castellanii mitochondria to oxidative stress

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The purpose of this study was to examine the effects of oxidative stress caused by hydroperoxide (H2O2) in the presence of iron ions (Fe2+) on mitochondria of the amoeba Acanthamoeba castellanii. We used isolated mitochondria of A. castellanii and exposed them to four levels of H2O2 concentration: 0.5, 5, 15, and 25 mM. We measured basic energetics of mitochondria: oxygen consumption in phosphorylation state (state 3) and resting state (state 4), respiratory coefficient rates (RC), ADP/O ratios, membrane potential (ΔΨm), ability to accumulate Ca2+ , and cytochrome c release. Our results show that the increasing concentrations of H2O2 stimulates respiration in states 3 and 4. The highest concentration of H2O2 caused a 3-fold increase in respiration in state 3 compared to the control. Respiratory coefficients and ADP/O ratios decreased with increasing stress conditions. Membrane potential significantly collapsed with increasing hydroperoxide concentration. The ability to accumulate Ca2+ also decreased with the increasing stress treatment. The lowest stress treatment (0.5 mM H2O2) significantly decreased oxygen consumption in state 3 and 4, RC, and membrane potential. The ADP/O ratio decreased significantly under 5 mM H2O2 treatment, while Ca2+ accumulation rate decreased significantly at 15 mM H2O2. We also observed cytochrome c release under increasing stress conditions. However, this release was not linear. These results indicate that as low as 0.5 mM H2O2 with Fe2+ damage the basic energetics of mitochondria of the unicellular eukaryotic organism Acanthamoeba castellanii.
Physical description
  • Institute of Dendrology, Polish Academy of Sciences, Kórnik, Poland
  • Adam Mickiewicz University, Institute of Molecular Biology and Biotechnology, Department of Bioenergetics, Poznań, Poland
  • Belzacq AS, Vieira HLA, Kroemer G, Brenner C (2002) The adenine nucleotide translocator in apoptosis. Biochimie 84: 167-176.
  • Bernardi P (1999) Mitochondrial transport of cations: channels, exchangers and permeability transition. Physiol Rev 79: 1127-1155.
  • Chen Q, Vazquez EJ, Moghaddas S, Hoppel CL, Lesnefsky EJ (2003) Production of reactive oxygen species by mitochondria: Central role of complex III. J Biol Chem 278: 36027-36031.
  • Czarna M, Jarmuszkiewicz W (2005) Activation of alternative oxidase and uncoupling protein lowers hydrogen peroxide formation in amoeba Acanthamoeba castellanii mitochondria. FEBS Lett 579: 3136-3140.
  • Domka-Popek A, Michejda JW (1986) The upake of Ca^2+ by mitochondria of amoeba supported by malate or ATP. Bull Soc Sci Lett 25.
  • Fleury C, Mignotte B, Vayssière JL (2002) Mitochondrial reactive oxygen species in cell death signaling. Biochimie 84: 131-141.
  • Garnier M, Dimchev A, Boujard N, Price J, Musto N, Papadopoulos V (1994) In vitro reconstitution of a functional benzodiazepine receptor from mouse Leyding tumor cells. Mol Pharmacol 45: 201-211.
  • Gincel D, Zaid H, Shoshan-Bermatz V (2001) Calcium binding and translocation by the voltage-dependent anion channel: a possible regulatory mechanism in mitochondrial function. Biochem J 358: 147-155.
  • Halestrap AP, McStay GP, Clarke SJ (2002) The permeability transition pore complex: another view. Biochimie 84: 153-166.
  • Jarmuszkiewicz W, Wagner AM, Wagner JM, Hryniewiecka L (1997) Immunological identification of the alternative oxidase of Acanthamoeba castellanii mitochondria. FEBS Lett 411: 110-114.
  • Kowaltowski A, Castilho R, Verces A (1996) Opening of the mitochondrial transition pore by uncoupling or inorganic phosphate in the presence of Ca^2+ is dependent on mitochondrial-generated reactive oxygen species. FEBS Lett 378: 150-152.
  • Kroemer G, Zamzani N, Susin SA (1997) Mitochondrial control of apoptosis. Immunol Today 18: 44-51.
  • Lenaz G, Bovina C, FormigginiG. Castelli-Parenti G (1999) Mitochondria, oxidative stress, and antioxidant defences. Acta Biochim Polon 46: 1-21.
  • Li Y, Trush MA (1998) Diphenyleneiodonium, an NAD(P)H oxidase inhibitor, also potently inhibits mitochondrial reactive oxygen species production. Biochem Biophys Res Commun 253: 295-299.
  • Martinou JC, Deshager S, Antonsson B (2000) Cytochrome c release from mitochondria: all or nothing. Nature Cell Biol 2: E41-E43.
  • Maxwell DP, Wang Y, McIntosh L (1999) The alternative oxidase lowers mitochondrial reactive oxygen production in plant cells. Proc Natl Acad Sci USA 96: 8271-8276.
  • Michejda JW, Schöneich J, Hryniewiecka L (1988) Activity of catalase in Acanthamoeba castellanii grown in conditions preferring the activity of cytochrome or alternative respiratory pathways. Bull Soc Sci Lett 26.
  • Minotti G, Aust SD (1987) The requirement for iron (III) in the initiation of lipid peroxidation by iron (II) and hydrogen peroxide. J Biol Chem 262: 1098-1104.
  • Papa S, Skulachev VP (1997) Reactive oxygen species, mitochondria, apoptosis and aging. Mol Cell Biochem 174: 305-319.
  • Purvis AC (1997) Role of the alternative oxidase in limiting superoxide production by plant mitochondria. Physiol Plant 100: 165-170.
  • Radi R, Bush KM, Freeman BA (1993) The role of cytochrome c and mitochondrial catalase in hydroperoxide-induced heart mitochondrial lipid peroxidation. Arch Biochem Biophys 300: 409-415.
  • Schein SJ, Colombini M, Finkelstein A (1976) Reconstitution in planar lipid bilayers of a voltage-dependent anion-selective channel obtained from Paramecium mitochondria. J Membr Biol 30: 99-120.
  • Shimizu S, Narita M, Tsujimoto Y (1999) Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature 399: 483-487.
  • Shimizu S, Ide T, Yanagida T, Tsujimoto Y (2000) Electrophysiological study of novel large pore formed by Bax and the voltage-dependent anion channel that is permeable to cytochrome c. J Biol Chem 275: 12321-12325.
  • Stridh H, Kimland M, Jones DP, Orrenius S, Hampton MB (1998) Cytochrome c release and caspase activation in hydrogen peroxide- and tributyltin-induced apoptosis. FEBS Lett 429: 351-355.
  • Sweetlove LJ, Heazlewood JL, Herald V, Holtzapffel R, Day DA, Leaver CJ, Millar AH (2002) The impact of oxidative stress on Arabidopsis mitochondria. Plant J 32: 891-904.
  • Ślesak I, Libik M, Karpińska B, Karpiński S, Miszalski Z (2007) The role of hydrogen peroxide in regulation of plant metabolism and cellular signalling in response to environmental stresses. Acta Biochim Polon 54: 39-50.
  • Tiwari BS, Belenghi B, Levine A (2002) Oxidative stress increased respiration and generation of reactive oxygen species, resulting in ATP depletion, opening of mitochondrial permeability transition, and programmed cell death. Plant Physiol 128: 1271-1281.
  • Vyssokikh M, Brdiczka D (2003) The function of complexes between the outer mitochondrial membrane pore (VDAC) and the adenine nucleotide translocase in regulation of energy metabolism and apoptosis. Acta Biochim Polon 50: 389-404.
  • Wudarczyk J, Dębska G, Lenartowicz E (1999) Zinc as inducer of the membrane permeability transition pore in rat liver mitochondria. Arch Biochem Biophys 363: 1-8.
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