EC Number   |
|---|
   1.1.3.4 | - |
| 1.1.3.4 | crystal structure analysis, PDB ID 1GPE |
| 1.1.3.4 | deglycosylation and purification to isoelectric homogeneity are important prerequisite steps to obtain crystals suitable for X-ray investigations. Crystals of the deglycosylated enzyme are reproducibly grown using ammonium sulfate as precipitant at pH 7.4 to 7.5. Crystals diffract to at least 2.0 A resolution and belong to the orthorhombic space group P2(1)2(1)2(1), with refined lattice constants of a = 59.3 A, b = 136.3 A and c = 156.7 A. Crystallized as a complex with the coenzyme FAD |
| 1.1.3.4 | native wild-type enzyme, hanging-drop vapor diffusion method at 4 °C by mixing equal volumes of 14 mg/mL AldO solution in 50 mM potassium phosphate buffer, pH 7.5, with reservoir solutions containing 0.1 M MES/HCl, pH 6.5, 0.2 M MgCl2 and 18-20% w/v PEG4000, 3-4 days, substrate incorporation by soaking the wild-type AldO crystals in a solution consisting of 0.1 M MES/HCl, pH 6.5, 0.2 M MgCl2, 25% w/v PEG 4000, 17.5% sucrose, and 25 mM substrate for 3 h, X-ray diffraction structure determination and analysis at 1.1-1.9 A resolution |
| 1.1.3.4 | of the partially deglycosylated enzyme, crystal structure determined by isomorphous replacement and refined to 2.3 A resolution |
| 1.1.3.4 | structure refined at 1.9 A resolution to an R value of 19.0% |
| 1.1.3.4 | tertiary structure determined by x-ray crystallography |
| 1.1.3.4 | the enzyme crystallizes from 1.3 M ammonium sulfate, 100 mM citrate/phosphate buffer pH 7.4 in the orthorhombic space group P212121, with unit-cell dimensions a = 57.6, b = 132.1, c = 151.3 A and one dimeric molecule per asymmetric unit. The structure is determined by molecular replacement and refined at 1.8 A resolution to an R value of 16.4% |
| 1.1.3.4 | vapor diffusion sitting drop method |
