Mitochondrial Reactive

Oxygen Species Alicia Kowaltowski

Mitochondrial Structure

Oxidative Phosphorylation

Cytochrome c Oxidase

Loschen G, Flohe L, Chance B. Respiratory chain linked H2O2 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

Nicholls and Ferguson, Bioenergetics 3, Academic Press, 2004

Complex I

Reverse Electron Transfer

Nicholls and Ferguson, Bioenergetics 3, Academic Press, 2004

Complex III

Nicholls and Ferguson, Bioenergetics 3, Academic Press, 2004

Turrens JF, J. Physiol. 2003, 552:335-344.

Electron Leakage at the Respiratory Chain

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

Dehydrogenases and Electron Leakage

H2O

O2.-

H2O2 HO.

2H+O2 .-

Fe2+ Fe3+

TPxcat

NOS

NO.Cad. Resp.

Inner Membrane

Outer Membrane

ONOO-

SOD

catalaseGPx

TPx

ROS Detected in Mitochondria

Mitochondrial NOS

Haynes et al., J. Bioenerg. Biomembr. 2004, 36:341-346.

L-arginine

L-citrullineNOS

NO.

- Alpha isoform of neuronal NOS

- Constitutive

- Regulates Complex IV

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

2O2-. + 2H+ H2O2 + O2

- Mutant MnSOD (-/-) animals do not survive unless treated with MnTBAP

Mitochondrial SODs

Green and O'Brien Biochim. Biophys. Acta. 1970, 197:31-39.

Levander et al. Biochem. Biophys. Res. Commun. 1974, 58:1047-1052.

H2O2 + 2GSH 2H2O + GSSGGPx

GSSG + NADPH 2GSH + NADP+Glut red.

NADP+ + NADH NADPH + NAD+Transhidr.

()

Glutathione Peroxidase/Reductase System

H2O2 + 2RSH 2H2O + RSSRTPx

RSSR + NADPH 2RSH + NADP+TR

Watabe et al. Eur. J. Biochem. 1997, 249:52-60.

2H2O2 2H2O + O2

Radi et al. J. Biol. Chem. 1991, 266:22028-22034.

Thioredoxin Peroxidase/Reductase System

Catalase

2Cyt c3+ + 2O2-. 2Cyt c2+ + 2O2

2Cyt c2+ + H2O2 + 2H+ 2Cyt c3+ + 2H2O

Cytochrome c

Skulachev FEBS Lett. 1998, 423:275-280.

Cyt c

I

II

IV

O2 + 4e- + 4H+ 2H2ONADH

SUCC

InnerMitochondrialMembrane

III

Electron Transport, OxPhos and ROS

+ + + + + + + + +++

H+ADP ATP

O2 + e- O2-. H2O2

Q

Loschen et al. FEBS Lett. 1971, 18:261-264. Korshunov et al. FEBS Lett. 1997, 416:15-18.

Mild Uncoupling Decreases ROS Release

Cyt c

I

II

IV

O2 + 4e- + 4H+ 2H2ONADH

SUCC

InnerMitochondrialMembrane

III

Electron Transport, OxPhos and ROS

+ + + + + + + + +++

H+ADP ATP

O2 + e- O2-. H2O2

Q

Negre-Salvayre et al. FASEB J. 1997, 11:809-815.

Uncoupling Proteins Regulate ROS Release

-

-

-

+

+

+

K+

- +

K+

H+

-

-

-

+

+

+

K+

ATP

MitoKATP Regulates ROS Release

cont

rol

ATP

ATP+DZXno

K

0.0

0.2

0.4

0.6

0.8

1.0

**

**

H2O

2 R

elea

se (

nmol

s . m

in-1 .

mg-1

)

Ferranti et al., 2003, FEBS Letts 536:51-5

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

Preconditioning and Postconditioning

wash ischemia reperfusion damage

IP damage

25 min 30 min 30 min

+ antioxidants damage

+ antioxidants

damage

Preconditioning and ROS

Vanden Hoek et al. J. Biol. Chem. 1998 273:18092-18098.

IP

Ischemic Protection

Mitochondrial ROS

mitoKATP UCP

Mild Uncoupling

ROS