11: The Reaction Mechanism of the Molybdenum Hydroxylase Xanthine Oxidoreductase: Evidence Against the Formation of Intermediates Having Metal-Carbon Bonds

ENDOR spectra of the catalytically relevant “very rapid” Mo(V) species generated in the course of the reaction of xanthine oxidoreductase with substrate have been examined by two different groups. While the data themselves are virtually identical, the analysis has been variously interpreted as supporting or refuting the existence of a molybdenum-carbon bond in the signal-giving species. While the basis for this difference in interpretation has now been generally agreed upon – the Mo-C distance in the signal-giving species is now understood to be too long to represent a direct Mo-C bond – independent information concerning the structure of the signal-giving species is highly desirable. Recently, several X-ray crystal structures of catalytically relevant complexes of the enzyme with several substrates and inhibitors have been reported. Taken together, these structures strongly and unambiguously support the interpretation that the intermediate giving rise to the “very rapid” EPR signal, as well as the Mo(IV) intermediate that precedes it in the reaction mechanism, has product coordinated to the active site molybdenum via the catalytically introduced hydroxyl group in a simple “end-on” fashion, with no metal-carbon bond character to the complex. The manner in which product is bound and its orientation within the active site provide important clues as to the specific catalytic roles of active sites in accelerating the reaction rate.

ENDOR spectra of the catalytically relevant “very rapid” Mo(V) species generated in the course of the reaction of xanthine oxidoreductase with substrate have been examined by two different groups. While the data themselves are virtually identical, the analysis has been variously interpreted as supporting or refuting the existence of a molybdenum-carbon bond in the signal-giving species. While the basis for this difference in interpretation has now been generally agreed upon – the Mo-C distance in the signal-giving species is now understood to be too long to represent a direct Mo-C bond – independent information concerning the structure of the signal-giving species is highly desirable. Recently, several X-ray crystal structures of catalytically relevant complexes of the enzyme with several substrates and inhibitors have been reported. Taken together, these structures strongly and unambiguously support the interpretation that the intermediate giving rise to the “very rapid” EPR signal, as well as the Mo(IV) intermediate that precedes it in the reaction mechanism, has product coordinated to the active site molybdenum via the catalytically introduced hydroxyl group in a simple “end-on” fashion, with no metal-carbon bond character to the complex. The manner in which product is bound and its orientation within the active site provide important clues as to the specific catalytic roles of active sites in accelerating the reaction rate.

ENDOR spectra of the catalytically relevant “very rapid” Mo(V) species generated in the course of the reaction of xanthine oxidoreductase with substrate have been examined by two different groups. While the data themselves are virtually identical, the analysis has been variously interpreted as supporting or refuting the existence of a molybdenum-carbon bond in the signal-giving species. While the basis for this difference in interpretation has now been generally agreed upon – the Mo-C distance in the signal-giving species is now understood to be too long to represent a direct Mo-C bond – independent information concerning the structure of the signal-giving species is highly desirable. Recently, several X-ray crystal structures of catalytically relevant complexes of the enzyme with several substrates and inhibitors have been reported. Taken together, these structures strongly and unambiguously support the interpretation that the intermediate giving rise to the “very rapid” EPR signal, as well as the Mo(IV) intermediate that precedes it in the reaction mechanism, has product coordinated to the active site molybdenum via the catalytically introduced hydroxyl group in a simple “end-on” fashion, with no metal-carbon bond character to the complex. The manner in which product is bound and its orientation within the active site provide important clues as to the specific catalytic roles of active sites in accelerating the reaction rate.

ENDOR spectra of the catalytically relevant “very rapid” Mo(V) species generated in the course of the reaction of xanthine oxidoreductase with substrate have been examined by two different groups. While the data themselves are virtually identical, the analysis has been variously interpreted as supporting or refuting the existence of a molybdenum-carbon bond in the signal-giving species. While the basis for this difference in interpretation has now been generally agreed upon – the Mo-C distance in the signal-giving species is now understood to be too long to represent a direct Mo-C bond – independent information concerning the structure of the signal-giving species is highly desirable. Recently, several X-ray crystal structures of catalytically relevant complexes of the enzyme with several substrates and inhibitors have been reported. Taken together, these structures strongly and unambiguously support the interpretation that the intermediate giving rise to the “very rapid” EPR signal, as well as the Mo(IV) intermediate that precedes it in the reaction mechanism, has product coordinated to the active site molybdenum via the catalytically introduced hydroxyl group in a simple “end-on” fashion, with no metal-carbon bond character to the complex. The manner in which product is bound and its orientation within the active site provide important clues as to the specific catalytic roles of active sites in accelerating the reaction rate.