EC Number   |
|---|
   1.17.3.2 | - |
| 1.17.3.2 | 2.25 A resolution, enzyme contains a molybdoptererin cofactor and two different [2Fe-2S] centers, enzyme is folded into four domains, the first two bind the iron sulfur centers, the last two are involved in molybtopterin binding |
| 1.17.3.2 | 2.5 A resolution, each enzyme subunit is composed of an N-terminal 20000 Da domain containing two iron sulfur centers, a central 40000 Da FAD domain and a C-terminal 85000 molybdopterin binding domain, the four redox centers are aligned in a linear fashion |
| 1.17.3.2 | batch method most suitable for crystallization |
| 1.17.3.2 | crystal structure determination and analysis of the enzyme in complex with a variety of substrates and substrate analogues, e.g. with 2-hydroxy-6-methylpurine or hypoxanthine, X-ray diffraction structure analysis at 1.8-3.1 A resolution |
| 1.17.3.2 | crystal structures for bovine xanthine oxidase in complex with indole-3-acetaldehyde and guanine, both at 1.6 A resolution is reported. In the case of indole-3-acetaldehyde, a modest, sterically allowed rotation juxtaposes the aldehyde group with the molybdenum center that allows hydroxylation to take place. With guanine, the substrate must rotate approximately 180° to present its 8 position to the active site molybdenum center for hydroxylation, and this rotation is sterically prohibited |
| 1.17.3.2 | enzme in complex with inhibitor quercetin, X-ray diffraction structure determintion and analysis at 2.0 A resolution |
| 1.17.3.2 | enzyme in complex with inhibitor 2-hydroxy-6-methylpurine, 10 mg/ml purified enzyme in 40 mM Tris-HCl, pH 7.8, 20 mM diphosphate, pH 8.5, 0.2 mM EDTA and 5 mM DTT, batch method mixing 0.02 ml of protein solution and 0.01 ml of precipitant solution, the latter containing of 12-14% PEG 8000, 0.1 M potassium phosphate, and 0.2 mM EDTA, pH 7.2, for 2-3 days at 25°C, X-ray diffraction structure determination and analysis at 2.3 A resolution, molecular replacement |
| 1.17.3.2 | purified enzyme in complex with hypoxanthine or 6-mercaptopurine, mixing of 0.01 ml of 5 mg/ml protein solution with 0.005-0.006 ml of 12% polyethylene glycol 8000 solution, at pH 7.0, crystallization in the darkness at 25°C, ligand binding by soaking of crystals, X-ray diffraction structure determination and analysis at 1.8 and 2.6 A resolution, respectively |
| 1.17.3.2 | purified recombinant C-terminally truncated mutant enzyme, crystals of the mutant protein are prepared in two ways: (a) crystallization of the protein directly after DTT treatment and (b) crystallization in the presence of DTT followed by extended soaks in mother liquor devoid of DTT to convert most of the protein to the XO form, X-ray diffraction structure determination and analysis at 2.0 A resolution. Comparisons of crystal structures of a stable wild-type XDH enzyme form, the triple mutant C535A/C992R/C1324, and the DELTAC truncated mutant XOR |
