   3.5.1.99 | recombinant fatty acid amide hydrolase mutant L192F/F194Y/A377T/S435N/I491V/V495M bound to carbamate inhibitor URB597 or inhibitor PF-3845, 25-30 mg/ml protein in 10 mM HEPES, pH 7.0, 500 mM NaCl, 2 mM dithiothreitol, and 0.08% n-undecyl-beta-D-maltoside or 0.1% n-decyl-beta-D-maltoside for inhibitor URB597-enzyme or PF-3845-enzyme conjugate, respectively, supplementation of the FAAHURB597 proteinsample with 1.6% benzyldimethyl(2-dodecyloxyethyl)-ammonium chloride and mixed at a 1:1 proportion with a crystallization buffer containing 30% PEG 400, 100 mM TrisHCl pH 7.5, and 100 mM MgCl2, supplementation of the FAAHPF-3845 protein sample with 1.6% benzyl-dimethyl-dodecyl ammonium bromide and mixed at a 1:1 ratio with a crystallization buffer containing 30% PEG 400, 100 mM 2-(N-morpholino)ethanesulfonate/NaOH, pH 5.5, and 400 mM LiCl, sitting drop vapor diffusion at 14°C, X-ray diffraction structure determination and analysis at 2.3 A resolution |
