   4.2.3.37 | purified wild-type enzyme and mutant F95N in complex with Mg2+, benzyl triethylammonium cation (BTAC), and diphosphate, sitting drop vapor diffusion method, mixing of 5 mm nl of 5 mg/ml protein in 300 mM NaCl, 20 mM Tris-HCl, pH 7.5, 10 mM MgCl2, 2 mM tris(2-carboxyethyl)phosphine (TCEP), 10% glycerol, 2 mM sodium pyrophosphate, and 2 mM BTAC with 600 nl of precipitant solution containing 0.17 M sodium acetate trihydrate, 85 mM sodium cacodylate trihydrate, pH 6.5, 25.5% PEG 8000, and 15% glycerol, addition of 100 nL of the 100fold dilution of wild-type enzyme crystallization seed stock, equilibration against a 0.1 ml reservoir of the precipitant solution at room temperature, 6 days, X-ray diffraction structure determination and analysis at 1.8 A and 2.51 A resolution, respectively |
744370 |
