Jezek 2013 Abstract MiP2013

Ježek P, Ježek J, Zelenka J, Dlasková A, Jabůrek M (2013) Antioxidant synergy of mitochondrial uncoupling protein UCP2 and phospholipase iPLA2γ. Mitochondr Physiol Network 18.08.

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Jezek P, Jezek J, Zelenka J, Dlaskova A, Jaburek M (2013)

Event: MiP2013

Recently, a modified mechanism [1] of originally suggested fatty acid cycling for mitochondrial uncoupling protein-1 (UCP1) confirmed all our original findings [2], including the necessity of fatty acid to approach UCP after its cleavage from membrane phospholipids [3]. We now tested a hypothesis that ubiquitous UCP2 provides feedback downregulation of oxidative stress in vivo via synergy with an H2O2–activated mitochondrial calcium–independent phospholipase A2 (mt-iPLA2), which cleaves off free fatty acids required for UCP2 as cycling substrates. We demonstrated UCP2 activation by free fatty acids (detected by gas chromatography/mass spectrometry) while observing that bovine serum albumin or GTP inhibit the tert-butylhydroperoxide– or H2O2–induced increase in respiration and decrease in membrane potential in lung and spleen mitochondria from control but not UCP2-knockout mice [4]. Upon induction of uncoupling, mitochondrial superoxide formation decreased instantly, but not when mt-iPLA2γwas blocked by R-bromoenol lactone (R-BEL). mt-iPLA2γ was alternatively activated by H2O2 produced in conjunction with the electron–transferring flavoprotein:ubiquinone oxidoreductase (ETFQOR), acting in fatty acid β-oxidation. We have also obtained similar findings in insulinoma INS-1E cells silenced for either UCP2 or iPLA2γ. Preliminary data showed that mt-iPLA2γis directly activated by H2O2. The evidence that this cytoprotective mechanism also exists in vivo stems from the observed increases in protein carbonylation in lung and spleen tissues and mitochondria, potentiated by R-BEL in wild-type but not in UCP2-knockout mice. We thus demonstrate for the first time a synergic antioxidant role between H2O2–activated mt-iPLA2γ and UCP2, because ablation of either mt-iPLA2γ or UCP2 prevented such a role. Our in vitro simulations also show for the first time that UCP2, functional due to fatty acids released by redox–activated mt-iPLA2γ, can suppress mitochondrial superoxide production by its uncoupling action.

H2O2–activated mt-iPLA2γ and UCP2 act in concert to protect against oxidative stress in vivo (cf. Figure).

• O2k-Network Lab: CZ Prague Jezek P

Labels: MiParea: Respiration 

Stress:RONS; Oxidative Stress"RONS; Oxidative Stress" is not in the list (Cell death, Cryopreservation, Ischemia-reperfusion, Permeability transition, Oxidative stress;RONS, Temperature, Hypoxia, Mitochondrial disease) of allowed values for the "Stress" property.  Organism: Mouse  Tissue;cell: Lung;gill, Islet Cell; Pancreas; Thymus"Islet Cell; Pancreas; Thymus" is not in the list (Heart, Skeletal muscle, Nervous system, Liver, Kidney, Lung;gill, Islet cell;pancreas;thymus, Endothelial;epithelial;mesothelial cell, Blood cells, Fat, ...) of allowed values for the "Tissue and cell" property.  Preparation: Intact cells, Isolated Mitochondria"Isolated Mitochondria" is not in the list (Intact organism, Intact organ, Permeabilized cells, Permeabilized tissue, Homogenate, Isolated mitochondria, SMP, Chloroplasts, Enzyme, Oxidase;biochemical oxidation, ...) of allowed values for the "Preparation" property.  Enzyme: Adenine Nucleotide Translocase"Adenine Nucleotide Translocase" is not in the list (Adenine nucleotide translocase, Complex I, Complex II;succinate dehydrogenase, Complex III, Complex IV;cytochrome c oxidase, Complex V;ATP synthase, Inner mt-membrane transporter, Marker enzyme, Supercomplex, TCA cycle and matrix dehydrogenases, ...) of allowed values for the "Enzyme" property., Uncoupling protein  Regulation: ATP production, Coupling efficiency;uncoupling, mt-Membrane potential, Redox state, Uncoupler, Fatty Acid"Fatty Acid" is not in the list (Aerobic glycolysis, ADP, ATP, ATP production, AMP, Calcium, Coupling efficiency;uncoupling, Cyt c, Flux control, Inhibitor, ...) of allowed values for the "Respiration and regulation" property.  Coupling state: LEAK, ROUTINE, OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property. 

HRR: Oxygraph-2k, Fluorometry"Fluorometry" is not in the list (Oxygraph-2k, TIP2k, O2k-Fluorometer, pH, NO, TPP, Ca, O2k-Spectrophotometer, O2k-Manual, O2k-Protocol, ...) of allowed values for the "Instrument and method" property. 

MiP2013 

Affiliations and author contributions

Institute of Physiology, Academy of Sciences, Prague, Czech Republic.

Email: [email protected]

Supported by GACR grants P302/10/0346 (PJ), P304/10/P204 (AD) and P303/11/P320 (JJ).

References

1. Fedorenko A, Lishko PV, Kirichok Y (2012) Mechanism of fatty-acid-dependent UCP1 uncoupling in brown fat mitochondria. Cell 151: 400-413.
2. Ježek P, Garlid KD (1990) New substrates and competitive inhibitors of the Cl- translocating pathway of the uncoupling protein of brown adipose tissue mitochondria. J Biol Chem 265: 19303-19311.
3. Jabůrek M, Miyamoto S, Di Mascio P, Garlid KD, Ježek P (2004) Hydroperoxy fatty acid cycling mediated by mitochondrial uncoupling protein UCP2. J Biol Chem 279: 53097–53102.
4. Jabůrek M, Ježek J, Zelenka J, Ježek P (2013) Antioxidant activity by a synergy of redox-sensitive mitochondrial phospholipase A2 and uncoupling protein-2 in lung and spleen. Int J Biochem Cell Biol 45: 816-825.