The conversion of xanthine dehydrogenase to xanthine oxidase and the role of the enzyme in reperfusion injury.
Although mammalian xanthine oxidase exists originally as a dehydrogenase form in freshly prepared samples, it is converted to an oxidase form during purification, either irreversibly by proteolysis or reversibly by sulfhydryl oxidation of the protein molecule. However, avoiding proteolysis the mammalian enzyme can be purified as an interconvertible form and thus can be used to compare directly the properties of xanthine dehydrogenase and the oxidase derived from the same enzyme molecule. The cDNAs encoding the enzyme have been cloned from several sources, and structural information is becoming available. The most significant difference between the two forms is the protein conformation around FAD, which changes the redox potential of the flavin and the reactivity of FAD with the electron acceptors, NAD and molecular oxygen. The flavin semiquinone is thermodynamically stable in xanthine dehydrogenase, but is unstable in xanthine oxidase. Detailed analyses by stopped-flow techniques suggest that the flavin semiquinone reacts with oxygen to form superoxide anion while the fully reduced flavin reacts to form hydrogen peroxide. Although xanthine dehydrogenase can produce greater amounts of superoxide anion than xanthine oxidase during xanthine-oxygen turnover, it seems to be physiologically insignificant because NAD inhibits almost completely the formation of superoxide anion. Although the involvement of this enzyme in reperfusion injury has been proposed, this seems to be more complex than originally envisaged and still remains to be established.
Nishino
T
T
Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo.
D000818:Animals; D015394:Molecular Structure; D015427:Reperfusion Injury / Q000201:enzymology*; D013329:Structure-Activity Relationship; D014968:Xanthine Dehydrogenase / Q000737:chemistry / Q000378:metabolism* / Q000502:physiology; D014969:Xanthine Oxidase / Q000378:metabolism*
D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase