EC Number   |
|---|
   3.1.1.72 | - |
| 3.1.1.72 | AcE forms a hexamer with a central substrate binding tunnel, and the intersubunit interactions are relatively weak. AcE also has a shorter loop and different residue composition in the beta4-alpha3 and beta5-alpha4 regions near the substrate binding site |
| 3.1.1.72 | catalytic machinery includes a catalytic triad (Ser119, His259, and Asp202) and an oxyanion hole (Cys40 and Ser120). Near the catalytic triad, aromatic residues Tyr39 and Trp160 form small pockets at both sides |
| 3.1.1.72 | fragment encoding the CE4 domain in complex with Cd2+, Mg2+ or Co2+, vapour-diffusion hanging-drop method |
| 3.1.1.72 | fragment encoding the CE4 domain, vapour-diffusion hanging-drop method |
| 3.1.1.72 | protein solution: ammonium sulfate in 50 mM citrate buffer, pH 5.3, X-ray diffraction structure determination at 0.9 and 1.1 A, analysis of the multiple conformations of the active site residues Ser90 and His187 |
| 3.1.1.72 | purified catalytic core of the enzyme at 10 mg/ml, hanging drop vapour diffusion method, room temperature, protein solution: sodium acetate, pH 5.0, reservoir solution: potassium/sodium tartrate 1.1 M, TES buffer 0.1 M, pH 8.2, in the ratio 2:1:1 with a Trixton X-100 solution, few weeks, X-ray diffraction structure determination and analysis at 1.9 A, structure model |
| 3.1.1.72 | purified recombinant enzyme free or in complex with paraoxon or diethyl phosphate, sparse matrix sitting drop vapour diffusion sampling procedure, 21°C, mixing of 0.0004 ml of protein solution, containing 8 mg/ml protein in 0.05 M Tris-HCl, pH 8.0, alone or supplemented with 0.5 mM paraoxon, with 0.0004 ml of reservoir solution containing 0.2 M MgCl2, 0.1 M HEPES, pH 7.5, and 30% w/v PEG 400, for enzyme in complex with diethyl phosphate, 0.2 M ammonium acetate, 0.1 M Bis-Tris, pH 5.5, and 45% v/v 2-methyl-2,4-pentanediol is used as the crystallization solution, X-ray diffraction structure determination and analysis at 1.9-2.7 A resolution, molecular replacement |
| 3.1.1.72 | purified recombinant enzyme mutant Y184F/W190P, hanging drop vapour diffusion method, mixing of 0.0025 ml of 3 mg/ml protein in 50 mM TrisHCl pH 7.0, 100 mM NaCl, and 0.02% NaN3, with 0.0025 ml of reservoir solution containing 16-21% PEG 3000, 0.1 M Tris-HCl, pH 7.5, and 200 mM calcium acetate, and equilibration over 0.5-1.0 ml reservoir solution, method optimization, X-ray diffraction structure determination and analysis at 2.3 A resolution |
| 3.1.1.72 | purified recombinant wild-type and selenomethionine-labeled enzyme, mixing of 0.0025 ml of 3-6 mg/ml protein solution with 0.0025 ml of reservoir solution, and equilibration against 1 ml of reservoir solution, 4 different conditions resulting in four different crystal forms, best conditions are obtained with 6 mg/ml protein and 1.2 M K tartrate, 0.3 M NaCl, 0.1 M imidazole buffer, pH 7.2, 1-3 days, X-ray diffraction structure determination and analysis at 1.70-1.85 A resolution, three-dimensional structure determination |
