【摘要】 旨在筛选及优化α-芋螺毒素与乙酰胆碱结合蛋白(ACh BPs)共结晶的条件,得到高分辨率的共结晶晶体。在昆虫表达系统中分泌表达海兔乙酰胆碱结合蛋白(Ac-ACh BP),并用凝胶色谱进行纯化,把纯化的乙酰胆碱结合蛋白与α-芋螺毒素GIC共结晶,利用结晶机器人及HAMPTON RESEARCH结晶试剂盒进行结晶及结晶条件的筛选与优化。结果显示,在0.6 mol/L Ammonium citrate dibasic,0.1 mol/L Sodium acetate trihydrate p H4.8的条件下生长的晶体较好,其分辨率可以达到2.1?。筛选与优化结晶条件(Ammonium citrate dibasic的摩尔浓度及0.1 mol/L Sodium acetate trihydrate的p H值)可以得到高分辨率的α-芋螺毒素GIC与Ac-ACh BP共结晶晶体。更多还原

【Abstract】 The aim is to screen and optimize the co-crystallization condition of α-conotoxin complex with acetylcholine bindingprotein(ACh BPs),and to obtain co-crystals with high resolution. ACh BP from Aplysia californica(Ac-ACh BP)was expressed in insectexpression system and purified by gel filtration chromatography. Then,the purified Ac-ACh BP was co-crystalized with α-conotoxin GIC,further,the HAMPTON RESEARCH crystal kit and the crystal robot were applied screening and optimization of the growth condition ofcrystallization. Results showed that crystal grew well under the growth condition of 0.6 mol/L ammonium citrate dibasic and p H4.8 of 0.1 mol/Lsodium acetate trihydrate,and the resolution of the crystal reached 2.1 ?. In conclusion,the co-crystal of α-conotoxin GIC complex withAc-ACh BP with a high resolution may be acquired via screening and optimization of crystallization condition(The molar concentration ofammonium citrate dibasic and the p H of 0.1 mol/L Sodium acetate trihydrate).更多还原