Reactions of oxidizing and reducing radical probes with lipoamide dehydrogenase.

One-electron redox radicals generated by radiation--chemical methods have been reacted with the oxidized (E) form of pig heart lipoamide dehydrogenase. The reducing radicals eaq- and CO2-. and O2-. do not measurably inactivate the enzyme, whereas the oxidizing species .OH and Br2-. do. The CO2-. anion forms the semiquinone radical .EH on the millisecond time scale, whereas at longer times only EH2 is observed. Evidence suggests that Br2-. oxidizes adjacent sulfhydryl groups to form a disulfide in a manner similar to the reaction of Cu2+ ions. With .OH, destruction of the flavin adenine dinucleotide (FAD) moiety is responsible for at least 50% of the enzyme inactivation. This destruction appears to be a result of secondary reactions which transfer damage from remote initial sites of attack to the flavin. Pathways for migration of eaq- damage also appear to exist.