EC Number   |
|---|
    2.5.1.17 | 3fold symmetric trimer of five counterclockwise helix bundles, one molecule in asymmetric unit, polypropylene glycol 400 molecule captured in putative active site (positively charged, important residues: Asp32, Arg118), crystals of selenomethionine derivative: space group P2(1)3, unit cell parameters: a, b, c: 84.67 A, hanging-drop vapour-diffusion method: protein solution (15 mg/ml) + reservoir solution (pH 8.1, 2.5 M ammonium sulphate, 2% polypropylene glycol 400) |
| 2.5.1.17 | apoenzyme and in complex with Mg2+/ATP, C-centred orthorhombic space group C222(1), unit cell parameters: a: 64.93 A, b: 137.08 A, c: 158.55 A, alpha, beta, gamma: 90°, one trimer in the asymmetric unit, sitting-drop combined with hanging-drop vapour-diffusion method: 13 mg/ml protein solution, precipitants: 1.55-1.6 M ammonium sulfate, 9-10% (v/v) dioxane (pH 6.5), for complex: 4 mM ATP and 4 mM Mg2+ |
| 2.5.1.17 | hanging-drop vapor diffusion method |
| 2.5.1.17 | mutants D35N (without tag) in complex with ATP and cob(II)alamin and R132K (without tag) in complex with ATP, thin plate crystals, space group P6(3), two monomers in the asymmetric unit, unit cell parameters: a, b: 65A, c: 169A, beta: 90°; vapour-diffusion under anoxic conditions, protein solution (15 mg/ml, containing ATP, hydroxycobalamin and a reducing system of NADH, FMN, and flavodoxin reductase), reservoir solution (incl. 14-16% PEG8000, pH6) |
| 2.5.1.17 | native (PDB: 2ZHY, with ordered N-terminal loop and formed active site) and in complex with ATP (PDB: 2ZHZ, substrate-binding cleft is widened and the N-terminal loop swung out and conformational shift of Arg129 side chain upon ATP-binding, similar structure to human and Lactobacillus reuteri PduO except for interaction of NH2 nitrogen atom of Arg11 with gamma-phosphate of ATP), five alpha helix bundle, monomeric subunits almost identical conformations in both structures, crystals: orthothrombic space group C222(1), three monomers in the asymmetric unit, unit cell parameters: a: 52.55/53.91, b: 148.98/148.17, c: 157.35/158.10, microcrystals (from sitting-drop vapour-diffusion at 22°C) scaled up by hanging-drop vapour diffusion, reservoir solution (pH5.7, 22% (w/v) isopropanol, 12% (v/v) PEG4000), PduO-MgATP complex: ATP soaked into native PduO crystals, molecular replacement using PDB: 2G2D as model |
| 2.5.1.17 | purified recombinant CobA in complex with ATP, four-coordinate cobalamin, and five-coordinate cobalamin, hanging drop vapour diffusion method, 0.002 ml of 10 mg/ml CobA protein in 20 mM Tris-HCl, pH 8.0, 20 mM NADH, 3 mM ATP, 4.5 mM MgCl2, and 2 mM HOCbl, is mixed with 0.002 ml of well solution containing 100 mM MES, pH 6.0, 320 mM NaCl, and 19.6% w/v PEG4000, X-ray diffraction structure determination and analysis at 1.95 A resolution |
| 2.5.1.17 | purified recombinant His-tagged enzyme in complex with its substrates, hanging drop vapour diffusion method, 20°C, 0.004 ml of 20 mg/ml protein in 10 mM Tris-HCl, pH 8.0, is mixed with 0.004 ml od precipitant solution containing 0.1 M HEPES, pH 8.5, 1.1 M ammonium sulfate, 2 mM ATP, 55 mM MgCl2, 165 mM NaCl, and 15 mM cob(I)alamin, 7 days, X-ray diffraction structure determination and analysis at 1.68 A resolution |
| 2.5.1.17 | purified recombinant PduO in complex with ATP, hanging drop vapor diffusion method, mixing o 0.001 ml of 13 mg/ml protein solution, with or without 4 mM ATP and 4 mM MgCl2, with 0.001 ml of optimized reservoir solution containing 100 mM MES, pH 6.5, 1.52 M ammonium sulfate, 9% v/v dioxane, followed by equilibration over 0.5 ml of the mother liquor, the cryoprotection solution contains 50 mM MES, pH 6.5, 0.76 M ammonium sulfate, 4.5% v/v dioxane, and 1.7 M sodium malonate, pH 7.0, X-ray diffraction structure determination and analysis |
| 2.5.1.17 | purified recombinant wild-type and selenomethionine-labeled enzymes, crystal growth from 0.4 M ammonium phosphate, 4% methyl-pentanediol, 5% glycerol, at 20°C, X-ray diffraction structure determination and analysis at 1.5-1.9 A resolution |
| 2.5.1.17 | sitting drop vapour diffusion method. The structure of PduOC co-crystallized with heme is solved (1.9 A resolution) showing an octameric assembly with four heme moieities |
