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

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

Abstract

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

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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