α-芋螺毒素GIC与Ac-AChBP共结晶条件的筛选与优化

doi:10.13560/j.cnki.biotech.bull.1985.2017.02.028

生物技术通报 ›› 2017, Vol. 33 ›› Issue (2): 192-198.doi: 10.13560/j.cnki.biotech.bull.1985.2017.02.028

α-芋螺毒素GIC与Ac-AChBP共结晶条件的筛选与优化

林波, 刘坤, 朱明月, 李伟, 董栩, 李孟森

海南医学院海南省肿瘤发生与干预重点实验室,海口 571199

收稿日期:2016-06-02 出版日期:2017-02-26 发布日期:2017-02-08
作者简介:林波,男,助理研究员,研究方向：蛋白质结构;E-mail：linbo_752@163.com
基金资助:
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）

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

LIN Bo, LIU Kun, ZHU Ming-yue, LI Wei, DONG Xu, LI Meng-sen

Hainan Provincial Key Laboratory of Carcinogenesis and Intervention,Hainan Medical College,Haikou 571199

Received:2016-06-02 Published:2017-02-26 Online:2017-02-08

摘要/Abstract

摘要： 旨在筛选及优化α-芋螺毒素与乙酰胆碱结合蛋白（AChBPs）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-AChBP）,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 pH4.8的条件下生长的晶体较好,其分辨率可以达到2.1 Å。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pH值）可以得到高分辨率的α-芋螺毒素GIC与Ac-AChBP共结晶晶体。

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

Abstract: 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-conotoxin 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 Å. 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）.

Key words: α-conotoxin GIC, acetylcholine-binding proteins, co-crystallization, optimization of crystal condition

林波, 刘坤, 朱明月, 李伟, 董栩, 李孟森. α-芋螺毒素GIC与Ac-AChBP共结晶条件的筛选与优化[J]. 生物技术通报, 2017, 33(2): 192-198.

LIN Bo, LIU Kun, ZHU Ming-yue, LI Wei, DONG Xu, LI Meng-sen. Screening and Optimization of Co-crystallization Condition of α-Conotoxin GIC Complex with Ac-AchBP[J]. Biotechnology Bulletin, 2017, 33(2): 192-198.

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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-AchBP

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