QM/MM studies of xanthine oxidase: variations of cofactor, substrate, and active-site Glu802.
In continuation of our previous QM/MM study on the reductive half-reaction of wild-type xanthine oxidase, we consider the effects of variations in the cofactor, the substrate, and the active-site Glu802 residue on the reaction mechanism. Replacement of the sulfido ligand in the natural cofactor by an oxo ligand leads to a substantial increase in the computed barriers, consistent with the experimentally observed inactivity of this modified cofactor, whereas the selenido form is predicted to have lower barriers and hence higher activity. For the substrate 2-oxo-6-methylpurine, the calculated pathways for three different tautomers show great similarity to those found previously for xanthine, contrary to claims in the literature that the mechanisms for these two substrates are different. Compared with the wild-type enzyme, the conversion of xanthine to uric acid follows a somewhat different pathway in the Glu802 --> Gln mutant which exhibits a lower overall activity, in agreement with recently published kinetic data. The present results confirm the basic stepwise reaction mechanism and the orientation of the substrate that has been proposed in our previous QM/MM work on aldehyde oxidoreductase and xanthine oxidase.
Metz
Sebastian
S
Max-Planck-Institut für Kohlenforschung, D-45470 Mülheim an der Ruhr, Germany.
Thiel
Walter
W
D020134:Catalytic Domain; D003067:Coenzymes / Q000378:metabolism*; D018698:Glutamic Acid; D008958:Models, Molecular; D016296:Mutagenesis; D009154:Mutation; D011687:Purines / Q000378:metabolism; D011789:Quantum Theory; D013816:Thermodynamics; D014969:Xanthine Oxidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*
D003067:Coenzymes; D011687:Purines; C071987:2-hydroxy-6-methylpurine; D018698:Glutamic Acid; D014969:Xanthine Oxidase