The molybdenum centers of xanthine oxidase and xanthine dehydrogenase. Determination of the spectral change associated with reduction from the Mo(VI) to the Mo(IV) state.
The UV-visible absorbance change associated with reduction of the molybdenum centers of xanthine oxidase and xanthine dehydrogenase has been determined using a double-difference technique. At pH 8.5, the Mo(VI) minus Mo(IV) difference spectrum seen with xanthine oxidase exhibits a positive feature at 420 nm, having an extinction change of approximately 3,000 M-1 cm-1 as well as evidence for a negative feature below 340 nm. In xanthine oxidase this change is found to exhibit a marked pH dependence, implicating protonation/deprotonation events associated with changes in the molybdenum oxidation state. Application of the double-difference protocol to the respective circular dichroism spectra of xanthine oxidase and xanthine dehydrogenase reveals appreciable CD changes at 420 and 580 nm associated with the reduction of the molybdenum center. The present results demonstrate a direct spectroscopic handle on the molybdenum centers of both xanthine oxidase and xanthine dehydrogenase.
Ryan
M G
MG
Department of Medical Biochemistry, Ohio State University, Columbus 43210, USA.
Ratnam
K
K
Hille
R
R
D000818:Animals; D002942:Circular Dichroism; D006863:Hydrogen-Ion Concentration; D008892:Milk / Q000201:enzymology; D008982:Molybdenum / Q000737:chemistry*; D010084:Oxidation-Reduction; D014968:Xanthine Dehydrogenase / Q000737:chemistry*; D014969:Xanthine Oxidase / Q000737:chemistry*
D008982:Molybdenum; D014968:Xanthine Dehydrogenase; D014969:Xanthine Oxidase