EC Number   |
|---|
   1.1.3.9 | ammonium sulfate precipitation, 1.7 A resolution |
| 1.1.3.9 | analysis of crystal structure, PDB ID 1GOF, of the processed (cofactor formed) Cu(II) site reveals the active site ligation. Residues His496, His581, cysteinylated-Tyr272, and an acetate ion form the four equatorial ligands. In the structure of Cu(II)-GO crystallized without acetate, PDB ID 1GOG, the fourth equatorial position is occupied by a weakly bound water. A second tyrosine, Tyr495, occupies an axial position with a Cu-O 2.62.7 A distance in these structures. Spectroscopical characterization of the geometric structure of the pre-processed Cu(I) active site using Cu K-edge X-ray absorption spectroscopy. Molecular modeling of the pre-processed Cu(I)-galactose oxidase active site and biogenesis reaction coordinate |
| 1.1.3.9 | crystal structures of the GAOV Cl2-Tyr272, and F2-Tyr272 incorporated variants at 1.48, 1.23, and 1.80 A resolution, respectively. Only one halogen atom remains in the cofactor. The structurally defined catalytic center with genetic unnatural tyrosine substitution is in the radical containing form as in the wild-type, i.e., Cu(II)-(Cl-Tyr radical-Cys) or Cu(II)-(F-Tyr radical-Cys). Thus an C-F/C-Cl bond cleavage is mediated by a mononuclear copper center |
| 1.1.3.9 | crystals of W290F and W290G are grown from protein as isolated, using hanging drop vapor diffusion from 1 to 2 M (NH4)2SO4 and 0.1 M sodium acetate at pH 4.0-5.0. To prepare the azide complex, wild-type galactose oxidase is dialyzed into 50 mM PIPES (pH 7.0) for 2 h, before addition of 20 mM sodium iethyl dithiocarbamate (DDC). The copper chelator DDC promotes the formation of orthorhombic crystals, which have more favorable crystal packing for substrate soaking experiments. The copper-free protein is crystallized in 13.5% PEG 8000, 200 mM calcium acetate, and 100 mM MES (pH 6.3) |
| 1.1.3.9 | hanging drop vapor diffusion, 1.7 A resolution |
| 1.1.3.9 | mutant C383S |
| 1.1.3.9 | sitting drop method. 2-5 mg/ml of enzyme mixed with equal volume of well solution. Crystals are soaked in oxygen-free Cu2+ solutions |
| 1.1.3.9 | sitting drop vapour diffusion, native, recombinant and mutant enzymes |
| 1.1.3.9 | wild-type enzyme and W290F mutant enzyme crystal structure analysis, PDB IDs 2EIE and 2EIC, respectively |
