F1F0-ATPase, Early Target of the Radical Initiator 2,2'-azobis-(2-amidinopropane) Dihydrochloride in Rat Liver Mitochondria in Vitro

Overview

Journal Biochem J

Specialty Biochemistry

Date 1996 Dec 1

PMID 8973568

Authors

F Beauseigneur

M Goubern

M F Chapey

J Gresti

C Vergely

M Tsoko

J Demarquoy

L Rochette

P Clouet

Affiliations

Soon will be listed here.

Abstract

This study was designed to determine which enzyme activities were first impaired in mitochondria exposed to 2,2'-azobis-(2-amidinopropane) dihydrochloride (AAPH), a known radical initiator. EPR spin-trapping revealed generation of reactive oxygen species although malondialdehyde formation remained very low. With increasing AAPH concentrations, State-3 respiration was progressively depressed with unaltered ADP/O ratios. A top-down approach demonstrated that alterations were located at the phosphorylation level. As shown by inhibitor titrations, ATP/ADP translocase activity was unaffected in the range of AAPH concentrations used. In contrast, AAPH appeared to exert a deleterious effect at the level of F1F0-ATPase, comparable with dicyclohexylcarbodi-imide, which alters Fo proton channel. A comparison of ATP hydrolase activity in uncoupled and broken mitochondria reinforced this finding. In spite of its pro-oxidant properties, AAPH was shown to act as a dose-dependent inhibitor of cyclosporin-sensitive permeability transition initiated by Ca2+, probably as a consequence of its effect on F1F0-ATPase. Resveratrol, a potent antiperoxidant, completely failed to prevent the decrease in State-3 respiration caused by AAPH. The data suggest that AAPH, when used under mild conditions, acted as a radical initiator and was capable of damaging F1F0-ATPase, thereby slowing respiratory chain activity and reducing mitochondrial antioxidant defences.

References

PENEFSKY H, CROSS R . Structure and mechanism of FoF1-type ATP synthases and ATPases. Adv Enzymol Relat Areas Mol Biol. 1991; 64:173-214. DOI: 10.1002/9780470123102.ch4. View

Groen A, Wanders R, Westerhoff H, van der Meer R, Tager J . Quantification of the contribution of various steps to the control of mitochondrial respiration. J Biol Chem. 1982; 257(6):2754-7. View

Bindoli A, Cavallini L, Jocelyn P . Mitochondrial lipid peroxidation by cumene hydroperoxide and its prevention by succinate. Biochim Biophys Acta. 1982; 681(3):496-503. DOI: 10.1016/0005-2728(82)90192-x. View

Marshansky V, Novgorodov S, Yaguzhinsky L . The role of lipid peroxidation in the induction of cation transport in rat liver mitochondria. The antioxidant effect of oligomycin and dicyclohexylcarbodiimide. FEBS Lett. 1983; 158(1):27-30. DOI: 10.1016/0014-5793(83)80669-3. View

Emanuel E, CARVER M, Solani G, Griffiths D . Differential inhibition of F0F1-ATPase-catalysed reactions in bovine-heart submitochondrial particles by organotin compounds. Biochim Biophys Acta. 1984; 766(1):209-14. DOI: 10.1016/0005-2728(84)90233-0. View

Cavallini L, Valente M, Bindoli A . Comparison of cumene hydroperoxide- and NADPH/Fe3+/ADP-induced lipid peroxidation in heart and liver submitochondrial particles. Mechanisms of protection by succinate. Biochim Biophys Acta. 1984; 795(3):466-72. DOI: 10.1016/0005-2760(84)90174-7. View

Rottenberg H . Membrane potential and surface potential in mitochondria: uptake and binding of lipophilic cations. J Membr Biol. 1984; 81(2):127-38. DOI: 10.1007/BF01868977. View

Beavis A, Brannan R, GARLID K . Swelling and contraction of the mitochondrial matrix. I. A structural interpretation of the relationship between light scattering and matrix volume. J Biol Chem. 1985; 260(25):13424-33. View

Smith P, Krohn R, Hermanson G, Mallia A, Gartner F, Provenzano M . Measurement of protein using bicinchoninic acid. Anal Biochem. 1985; 150(1):76-85. DOI: 10.1016/0003-2697(85)90442-7. View

10.

Yamamoto Y, Niki E, Kamiya Y, Miki M, Tamai H, Mino M . Free radical chain oxidation and hemolysis of erythrocytes by molecular oxygen and their inhibition by vitamin E. J Nutr Sci Vitaminol (Tokyo). 1986; 32(5):475-9. DOI: 10.3177/jnsv.32.475. View

11.

Terao K, Niki E . Damage to biological tissues induced by radical initiator 2,2'-azobis(2-amidinopropane) dihydrochloride and its inhibition by chain-breaking antioxidants. J Free Radic Biol Med. 1986; 2(3):193-201. DOI: 10.1016/s0748-5514(86)80070-8. View

12.

Brand M, Murphy M . Control of electron flux through the respiratory chain in mitochondria and cells. Biol Rev Camb Philos Soc. 1987; 62(2):141-93. DOI: 10.1111/j.1469-185x.1987.tb01265.x. View

13.

Niki E . Antioxidants in relation to lipid peroxidation. Chem Phys Lipids. 1987; 44(2-4):227-53. DOI: 10.1016/0009-3084(87)90052-1. View

14.

Buettner G . Spin trapping: ESR parameters of spin adducts. Free Radic Biol Med. 1987; 3(4):259-303. DOI: 10.1016/s0891-5849(87)80033-3. View

15.

Hafner R, Brown G, Brand M . Thyroid-hormone control of state-3 respiration in isolated rat liver mitochondria. Biochem J. 1990; 265(3):731-4. PMC: 1133694. DOI: 10.1042/bj2650731. View

16.

Hafner R, Brown G, Brand M . Analysis of the control of respiration rate, phosphorylation rate, proton leak rate and protonmotive force in isolated mitochondria using the 'top-down' approach of metabolic control theory. Eur J Biochem. 1990; 188(2):313-9. DOI: 10.1111/j.1432-1033.1990.tb15405.x. View

17.

GUNTER T, Pfeiffer D . Mechanisms by which mitochondria transport calcium. Am J Physiol. 1990; 258(5 Pt 1):C755-86. DOI: 10.1152/ajpcell.1990.258.5.C755. View

18.

Lissi E, Clavero N . Inactivation of lysozyme by alkylperoxyl radicals. Free Radic Res Commun. 1990; 10(3):177-84. DOI: 10.3109/10715769009149886. View

19.

Brand M, DAlessandri L, Reis H, Hafner R . Stimulation of the electron transport chain in mitochondria isolated from rats treated with mannoheptulose or glucagon. Arch Biochem Biophys. 1990; 283(2):278-84. DOI: 10.1016/0003-9861(90)90643-d. View

20.

Nishimura M, Ito T, Chance B . Studies on bacterial photophosphorylation. III. A sensitive and rapid method of determination of photophosphorylation. Biochim Biophys Acta. 1962; 59:177-82. View