  3.1.1.56 | apo-enzyme (PDB: 1PV1) and mutant W197I (PDB: 3C6B) following inhibition through phosphorylation with diethyl p-nitrophenyl phosphate, sitting drop: pH 4.6, 25.5% (w/v) polyethylene glycol (PEG) 4000, 15% glycerol, 17°C, crystals: space group: P2(1)2(1)2(1) for PDB: 1PV1 (four molecules in asymmetric unit, dimer of dimers), C2 for PDB: 3C6B, structure: PDB: 1PV1: three-layer sandwich without disulfide bonds, catalytic triad: S161, H276, D241 and adjacent C60, oxyanion hole: L58 and M162, PDB: 3C6B: tetrahedral shape, sulfenic acid at residue C60, structural superimposition of wild-type and mutant W197I: natural resistance to paraoxon (diethyl p-nitrophenyl phosphate, an organophosphorus inhibitor) due to steric hindrance by W197 within the enzymes acyl pocket (L58, W102, I189, F243, L248, W197) |
| 3.1.1.56 | hanging drop: 16°C, 3 days, reservoir solution: sodium citrate pH 5.6, 200 mM ammonium acetate, 30% (w/v) polyethylene glycol (PEG) 4000, crystal: space group P2(1), unit cell parameters: a: 51.54, b: 70.72, c: 65.01, alpha: 90°, beta: 108.84°, gamma: 90°, 2 molecules in the asymmetric unit, molecular replacement using PDB: 1PV1, co-crystallization and soaking with substrate failed, structure: several insertions in canonical alpha/beta-hydrolase fold, GxSxG motif, active site in positively charged cleft including residues S149, D226, and H260, and aromatic residues (H148, F172 W183, F228, H260, Y262) lined at the surface, catalysis mediated by residues S153, H264, and D230, oxyanion hole formed by e.g. L54 and M150 |
