Reactive oxygen species-mediated inactivation of pyruvate dehydrogenase.
Brain ischemia reperfusion causes increased formation of reactive oxygen species (ROS). Activity of the mitochondrial enzyme pyruvate dehydrogenase (PDH) has been shown to undergo a significant decrease following reperfusion of the ischemic tissue. We have examined the effect of a superoxide radical-generating system (xanthine oxidase/hypoxanthine, XO/HX) on the activity of this enzyme. Incubation of PDH in the presence of XO/HX resulted in its inactivation. The degree of the inactivation was dependent on the amount of XO present, which correlated linearly with the concentration of superoxide radical generated by this system. The activity of lactate dehydrogenase, an enzyme resistant to inactivation by ischemia reperfusion, was not affected by this system. Superoxide dismutase partially prevented and catalase exerted a nearly complete protective effect against the inactivation of PDH. Deferoxamine was partially protective. The sulfhydryl protective reagents, dithiothreitol and glutathione, prevented the inactivation of PDH, even though to varying degrees, which implicates sulfhydryl oxidation. A hydroxyl radical-generating system (hydrogen peroxide irradiated with ultraviolet radiation) effectively inactivated PDH. These results demonstrate that PDH is susceptible to damage and inactivation by ROS and point to the involvement of Fenton chemistry and hydroxyl radicals formed through it in PDH inactivation by XO/HX. A similar mechanism may be responsible for the PDH inactivation during ischemia/reperfusion.
Tabatabaie
T
T
Free Radical Biology and Aging Research Program, Oklahoma Medical Research Foundation, 825 N.E. 13th Street, Oklahoma City, Oklahoma, 73104, USA.
Potts
J D
JD
Floyd
R A
RA
D000818:Animals; D002374:Catalase / Q000378:metabolism; D002417:Cattle; D003676:Deferoxamine / Q000494:pharmacology; D004229:Dithiothreitol / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000378:metabolism / Q000494:pharmacology*; D005978:Glutathione / Q000494:pharmacology; D006861:Hydrogen Peroxide / Q000494:pharmacology; D019271:Hypoxanthine / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008353:Mannitol / Q000494:pharmacology; D010663:Phenylmercury Compounds / Q000494:pharmacology; D011768:Pyruvate Dehydrogenase Complex / Q000037:antagonists & inhibitors*; D017382:Reactive Oxygen Species / Q000378:metabolism; D013482:Superoxide Dismutase / Q000378:metabolism; D013552:Swine; D014969:Xanthine Oxidase / Q000378:metabolism*
D004791:Enzyme Inhibitors; D010663:Phenylmercury Compounds; D011768:Pyruvate Dehydrogenase Complex; D017382:Reactive Oxygen Species; C002910:4-hydroxymercuribenzenesulfonate; D019271:Hypoxanthine; D008353:Mannitol; D006861:Hydrogen Peroxide; D007770:L-Lactate Dehydrogenase; D002374:Catalase; D013482:Superoxide Dismutase; D014969:Xanthine Oxidase; D005978:Glutathione; D003676:Deferoxamine; D004229:Dithiothreitol