温度: 293 K / 手法: 蒸気拡散法, シッティングドロップ法 / pH: 7 詳細: Directly after size exclusion chromatography chMDA5 deltaCARD-Q was mixed with 10 bp dsRNA in a 1:1 molar ratio and incubated for 30 minutes on ice. The complex was concentrated using an ...詳細: Directly after size exclusion chromatography chMDA5 deltaCARD-Q was mixed with 10 bp dsRNA in a 1:1 molar ratio and incubated for 30 minutes on ice. The complex was concentrated using an Amicon Ultra concentrator to around 10 mg/ml and 2 mM AMPPNP (adenosine 5prime-(beta,gamma-imido)triphosphate lithium salt hydrate) was added. Sample and reservoir buffer (0.025 M Bis-Tris pH 6.5, 0.075 M succinic acid pH 7.0, 12-14% PEG 3350, 2% sucrose) were mixed in a 2:1 ratio. Three hours after setup, cover glasses with drops were transferred from a reservoir containing 12-14% PEG 3350 to one containing 8% PEG 3350. Crystals grew in one week at 20 C and were harvested in cryo-protectant solution (0.025 M Bis-Tris pH 6.5, 0.075 M succinic acid pH 7.0, 25% 3350, 10% ethylene glycol) before flash freezing with liquid nitrogen.
解像度: 2.6→50 Å / Cor.coef. Fo:Fc: 0.889 / Cor.coef. Fo:Fc free: 0.875 / SU B: 13.362 / SU ML: 0.294 / 交差検証法: THROUGHOUT / ESU R: 1.056 / ESU R Free: 0.363 / 詳細: HYDROGENS HAVE BEEN ADDED IN THE RIDING POSITIONS