Mechanism of transition from xanthine dehydrogenase to xanthine oxidase: effect of guanidine-HCL or urea on the activity

Abstract

Mammalian xanthine oxidoreductase can be converted from the dehydrogenase to the oxidase form, either reversibly by formation of disulfide bridges or irreversibly by proteolytic cleavage within the xanthine oxidoreductase protein molecule. A tightly packed amino acid cluster stabilizes the dehydrogenase form, and disruption of this cluster is accompanied with rearrangement of the active site loop. Here, we show that the conversion occurs in the presence of guanidine-HCl or urea. We propose that xanthine dehydrogenase and oxidase are in a thermodynamic equilibrium that can be shifted by disruption of the amino acid cluster with a denaturant.