Effects of photodynamic action on respiration in nonphosphorylating mitochondria.
We have studied the effects of singlet oxygen produced by photodynamic action on respiration in nonphosphorylating mitochondria (state 4). Isolated rat liver mitochondria were incubated with 3 microM hematoporphyrin and irradiated at 365 nm with a fluence rate of 25 W/m2. After short durations of irradiation, state 4 respiration with beta-hydroxybutyrate as substrate increases while respiration with succinate is negligibly affected. When mitochondria have been uncoupled with carbonylcyanide-p-trifluoromethoxyphenyl hydrazone before irradiation, no change occurs in beta-hydroxybutyrate-driven respiration, while succinate-driven respiration strongly decreases. Stimulation of state 4 NADH respiration cannot be explained by slippage of the NADH ubiquinone oxidoreductase because the stoichiometry of the redox pump was found insensitive to photodynamic action. In the light of the metabolite theory for linear enzymatic chains applied to state 4 respiration (Brand et al., Biochem. J. 255, 535-539, 1988), these results suggest that stimulation of NADH respiration is simply due to an increase of membrane leaks which occurs after irradiation. In the case of succinate-driven respiration, a strong inhibition of succinate dehydrogenase activity has been demonstrated after irradiation. It can be suggested that this inhibition introduces a negative control coefficient over state 4 respiration, counterbalancing the effects due to leakage.
INSERM U 201 et CNRS URA 481, Muséum National d'Histoire Naturelle, 43 rue Cuvier, Paris Cédex 05, 75231, France.
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Salet
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Moreno
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Ricchelli
D000818:Animals; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000528:radiation effects; D019069:Cell Respiration / Q000187:drug effects / Q000528:radiation effects*; D003470:Culture Media / Q000378:metabolism; D004533:Egtazic Acid / Q000494:pharmacology; D004579:Electron Transport / Q000187:drug effects / Q000528:radiation effects; D006415:Hematoporphyrins / Q000494:pharmacology; D007425:Intracellular Membranes / Q000187:drug effects / Q000528:radiation effects; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism* / Q000528:radiation effects*; D009840:Oligomycins / Q000494:pharmacology; D010084:Oxidation-Reduction; D010766:Phosphorylation / Q000528:radiation effects; D010777:Photochemistry; D017494:Proton Pumps / Q000187:drug effects / Q000528:radiation effects; D051381:Rats; D014466:Ultraviolet Rays
D003470:Culture Media; D006415:Hematoporphyrins; D009840:Oligomycins; D017494:Proton Pumps; D004533:Egtazic Acid