Singlet oxygen produced by photodynamic action causes inactivation of the mitochondrial permeability transition pore.
We have studied the effects of singlet oxygen produced by photodynamic action on the cyclosporin A-sensitive permeability transition (PT) in isolated rat liver mitochondria. Mitochondria were incubated with 3 microM hematoporphyrin and irradiated at 365 nm with a fluence rate of 25 watts/m2. For short durations of irradiation (60 s) the adenine nucleotide translocase was inactivated, but mitochondria retained their ability to form a proton electrochemical gradient and accumulated Ca2+ and Pi at the same rate as non-irradiated controls. Strikingly, however, the oxidative effects of photodynamic action prevented opening of the PT pore which is normally induced by Ca2+ plus Pi or by treatment with diethyl pyrocarbonate (a histidine reagent) or diamide (a thiol oxidant). We show that the most likely targets for photodynamic action are critical histidines that undergo degradation. Irradiated, hematoporphyrin-loaded mitochondria treated with diethyl pyrocarbonate or diamide still undergo the PT when treated with phenylarsine oxide, which reacts with a critical dithiol involved in pore modulation (Petronilli, V., Costantini, P., Scorrano, L., Colonna, R., Passamonti, S., and Bernardi, P. (1994) J. Biol. Chem. 269, 16638-16642). These data suggest (i) that the dithiol cysteines are not oxidized by photodynamic action, but rather became inaccessible to oxidants; and (ii) that irradiation of hematoporphyrin-loaded mitochondria does not lead to pore denaturation, but rather to site-selective inactivation of discrete pore functional domains.
Salet
C
C
Laboratoires de Biophysique et de Photobiologie, INSERM U 201 et CNRS URA 481, Muséum National d'Histoire Naturelle, 43 rue Cuvier, 75231 Paris Cédex 05, France.
Moreno
G
G
Ricchelli
F
F
Bernardi
P
P
D000818:Animals; D002118:Calcium / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D003958:Diamide / Q000494:pharmacology; D004047:Diethyl Pyrocarbonate / Q000494:pharmacology; D007473:Ion Channels / Q000037:antagonists & inhibitors*; D008565:Membrane Proteins / Q000037:antagonists & inhibitors*; D008930:Mitochondria, Liver / Q000187:drug effects / Q000378:metabolism*; D010100:Oxygen / Q000378:metabolism*; D010101:Oxygen Consumption / Q000187:drug effects; D010777:Photochemistry; D018272:Porins; D051381:Rats; D026082:Singlet Oxygen; D050994:Voltage-Dependent Anion Channels
D007473:Ion Channels; D008565:Membrane Proteins; D018272:Porins; D050994:Voltage-Dependent Anion Channels; D003958:Diamide; D026082:Singlet Oxygen; D016572:Cyclosporine; D004047:Diethyl Pyrocarbonate; D010100:Oxygen; D002118:Calcium