α-芋螺毒素GIC与Ac-AChBP共结晶条件的筛选与优化  

Screening and Optimization of Co-crystallization Condition of α-Conotoxin GIC Complex with Ac-Ach BP

作　　者：林波[1] 刘坤[1] 朱明月[1] 李伟[1] 董栩[1] 李孟森[1] 

机构地区：[1]海南医学院海南省肿瘤发生与干预重点实验室,海口571199

出　　处：《生物技术通报》2017年第2期192-198,共7页Biotechnology Bulletin

基　　金：国家自然科学基金项目(31560243;81560450);海南医学院科研培育基金项目(HY2013-05)

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。The aim is to screen and optimize the co-crystallization condition of α-conotoxin complex with acetylcholine binding protein （ AChBPs ）, and to obtain co-crystals with high resolution. AChBP from Aplysia californica （Ac-AChBP） was expressed in insect expression system and purified by gel filtration chromatography. Then, the purified Ac-AChBP was co-crystalized with α-eonotoxin GIC, further, the HAMPTON RESEARCH crystal kit and the crystal robot were applied screening and optimization of the growth condition of crystallization. Results showed that crystal grew well under the growth condition of 0.6 mol/L ammonium citrate dibasic and pH4.8 of 0.1 mol/L sodium acetate trihydrate, and the resolution of the crystal reached 2.1 A. In conclusion, the co-crystal of α-conotoxin GIC complex with Ac-AChBP with a high resolution may be acquired via screening and optimization of crystallization condition （ The molar concentration of ammonium citrate dibasic and the pH of 0.1 mol/L Sodium acetate trihydrate ） .

关 键 词：α-芋螺毒素GIC 乙酰胆碱结合蛋白 共结晶 结晶条件优化 

分 类 号：Q51[生物学—生物化学]