1. Mitochondrial Reactive Oxygen Species Alicia Kowaltowski

2. Mitochondrial Structure

3. Oxidative Phosphorylation

4. Cytochrome c Oxidase

5. Loschen G, Flohe L, Chance B. Respiratory chain linked H 2 O 2 production in pigeon heart mitochondria. FEBS Lett., 1971 18:261-264. (Effects of different respiratory states) Boveris A, Chance B. The mitochondrial generation of hydrogen peroxide. General properties and effect of hyperbaric oxygen. Biochem. J., 1973 134:707-716. (Effects of respiratory inhibitors rotenone and antimycin) Dionisi O, Galeotti T, Terranova T, Azzi A. Superoxide radicals and hydrogen peroxide formation in mitochondria from normal and neoplastic tissues. Biochim. Biophys. Acta 1975 403:292-300. (Detection of superoxide radicals) Mitochondrial Reactive Oxygen Species Formation

6. Nicholls and Ferguson, Bioenergetics 3, Academic Press, 2004 Complex I

7. Reverse Electron Transfer Nicholls and Ferguson, Bioenergetics 3, Academic Press, 2004

8. Complex III Nicholls and Ferguson, Bioenergetics 3, Academic Press, 2004

9. Turrens JF, J. Physiol. 2003, 552:335-344. Electron Leakage at the Respiratory Chain

10. Tretter and Adam-Vizi J. Neurosci. 2004, 24:7771-7778. Starkov et al. J. Neurosci. 2004, 24:7779-7788. Dehydrogenases and Electron Leakage

11. H2OH2O O 2.- H2O2H2O2 HO. 2H + O 2.- Fe 2+ Fe 3+ TPxcat NOS NO. Cad. Resp. Inner Membrane Outer Membrane ONOO - SOD catalase GPx TPx ROS Detected in Mitochondria

12. Mitochondrial NOS Haynes et al., J. Bioenerg. Biomembr. 2004, 36:341-346. L-arginine L-citrulline NOS NO. - Alpha isoform of neuronal NOS - Constitutive - Regulates Complex IV

13. Weisiger and Fridovich J. Biol. Chem. 1973, 248:4793-4796. Sturtz et al J. Biol. Chem. 2001, 276:38084-38089. Okado-Matsumoto and Fridovich J. Biol. Chem. 2001, 276:38388-383893 - MnSOD in the matrix, CuZnSOD in the intermembrane space 2O 2 -. + 2H + H 2 O 2 + O 2 - Mutant MnSOD (-/-) animals do not survive unless treated with MnTBAP Mitochondrial SODs

14. Green and O'Brien Biochim. Biophys. Acta. 1970, 197:31-39. Levander et al. Biochem. Biophys. Res. Commun. 1974, 58:1047-1052. H 2 O 2 + 2GSH 2H 2 O + GSSG GPx GSSG + NADPH 2GSH + NADP + Glut red. NADP + + NADH NADPH + NAD + Transhidr. (  ) Glutathione Peroxidase/Reductase System

15. H 2 O 2 + 2RSH2H 2 O + RSSR TPx RSSR + NADPH2RSH + NADP + TR Watabe et al. Eur. J. Biochem. 1997, 249:52-60. 2H 2 O 2 2H 2 O + O 2 Radi et al. J. Biol. Chem. 1991, 266:22028-22034. Thioredoxin Peroxidase/Reductase System Catalase

16. 2Cyt c 3+ + 2O 2 -. 2Cyt c 2+ + 2O 2 2Cyt c 2+ + H 2 O 2 + 2H + 2Cyt c 3+ + 2H 2 O Cytochrome c Skulachev FEBS Lett. 1998, 423:275-280.

17. Cyt c I II IV O 2 + 4e - + 4H + 2H 2 O NADH SUCC Inner Mitochondrial Membrane III Electron Transport, OxPhos and ROS +++ +++++ +++ H+H+ ADP ATP O 2 + e - O 2 -. H 2 O 2 Q

18. Loschen et al. FEBS Lett. 1971, 18:261-264. Korshunov et al. FEBS Lett. 1997, 416:15-18. Mild Uncoupling Decreases ROS Release

19. Cyt c I II IV O 2 + 4e - + 4H + 2H 2 O NADH SUCC Inner Mitochondrial Membrane III Electron Transport, OxPhos and ROS +++ +++++ +++ H+H+ ADP ATP O 2 + e - O 2 -. H 2 O 2 Q

20. Negre-Salvayre et al. FASEB J. 1997, 11:809-815. Uncoupling Proteins Regulate ROS Release

21. - - - + + + K+K+ -+ K+K+ H+H+ - - - + + + K+K+ ATP MitoK ATP Regulates ROS Release Ferranti et al., 2003, FEBS Letts 536:51-5

22. Mattiasson G, Shamloo M, Gido G, Mathi K, Tomasevic G, Yi S, Warden CH, Castilho RF, Melcher T, Gonzalez-Zulueta M, Nikolich K, Wieloch T. Uncoupling protein-2 prevents neuronal death and diminishes brain dysfunction after stroke and brain trauma. Nat. Med. 2003, 9:1062-1068. Hoerter J, Gonzalez-Barroso MD, Couplan E, Mateo P, Gelly C, Cassard-Doulcier AM, Diolez P, Bouillaud F. Mitochondrial uncoupling protein 1 expressed in the heart of transgenic mice protects against ischemic- reperfusion damage. Circulation 2004, 110:528-533. Holmuhamedov EL, Jahangir A, Oberlin A, Komarov A, Colombini M, Terzic A. Potassium channel openers are uncoupling protonophores: implication in cardioprotection. FEBS Lett. 2004, 568:167-170. Barros MH, Bandy B, Tahara EB, Kowaltowski AJ. Higher respiratory activity decreases mitochondrial reactive oxygen release and increases life span in Saccharomyces cerevisiae. J Biol Chem. 2004, 279:49883-49888. Tissue Protection Through Uncoupling

23. Preconditioning and Postconditioning washischemiareperfusion  damage IP  damage 25 min30 min + antioxidants  damage + antioxidants  damage

24. Preconditioning and ROS Vanden Hoek et al. J. Biol. Chem. 1998 273:18092-18098.

25. IP Ischemic Protection  Mitochondrial ROS mitoK ATP UCP Mild Uncoupling  ROS