EC Number   |
|---|
    5.3.99.2 | - |
| 5.3.99.2 | crystals of H-PGDS are first grown by hanging-drop vapor diffusion method with PEG6000 as the precipitant at 20°C in the presence of 5 mM Mg2+ and the native crystals are then soaked in the precipitant solution with a saturating concentration of 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazyl) tetrazolium chloride for 2 weeks. Structure of the enzyme complexed with 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazyl) tetrazolium chloride at 1.9 A resolution in the presence of Mg2+. The styryl group of the inhibitor penetrates to the bottom of the active site cleft, and the tetrazole ring is stabilized by the stacking interaction with TRp104, inducing large movement around the alpha5-helix, which causes the space group of the complex crystal to change from P2(1) to P1 upon binding of 2-(2'-benzothiazolyl)-5-styryl-3-(4'-phthalhydrazyl) tetrazolium chloride |
| 5.3.99.2 | H-PGDS is crystallized in a microgravity environment at 20°C, using 30% (w/v) PEG 6000, 10 mM dithiothreitol, 10 mM glutathione, 1% (v/v) dioxane and 1 mM magnesium chloride in 50 mM Tris-HCl pH 8.4 |
| 5.3.99.2 | hanging drop vapor diffusion method, using 0.1 M MES/NaOH at pH 6.5, 0.005 M MgCl2 and 24% (w/v) PEG 3350 |
| 5.3.99.2 | in complex with glutathione, sitting drop vapor diffusion method, using 0.1 M Tris-HCl, pH 8.0, 28% (w/v) PEG4000 |
| 5.3.99.2 | macro-seeding using a solution containing 14% polyethylene glycol 6000, 50 mM Tris-HCl, pH 8.4, 5 mM reduced glutathione, 5 mM dithiothreitol, 2.5 mM CaCl2 or MgCl2 and 1% dioxane, structure resolution at 1.8 A |
| 5.3.99.2 | mutant C65A in complex with oleic acid and palmitoleic acid, hanging drop vapor diffusion method, using 0.1 M citric acid (pH 4.0) and 1.7 M ammonium sulfate |
| 5.3.99.2 | mutant DELTA1-24_C65A L-PGDS is crystallized in two different crystal forms representing the conformational change between the open and closed states of the cavity of the beta-barrel, the structures are determined to resolutions of 2.1 and 2.0 A |
| 5.3.99.2 | mutant enzymes C65A/K59A, C65A, C65A/W54F, C65A/W112F and C65A/W54F/W112F hanging drop vapor diffusion method, using 0.2 M ammonium sulfate, 30% (w/v) PEG 4000 |
| 5.3.99.2 | NMR solution structure, enzyme consists of an eight-stranded, antiparallel beta-barrel and a long alpha-helix associated with the outer surface of the barrel. The interior of the barrel forms a hydrophobic cavity containing two pockets. Prostaglandin H2 almost fully occupies hydrophilic pocket 1, in which C65 is located, and all-trans retinoic acid occupies hydrophilic pocket 2 |
