人源雄激素受体DNA结合结构域的原核表达与其响应元件的复合物结晶

Prokaryotic expression of homo androgen receptor DNA binding domain and co-crystallization with androgen response element

摘要:

为获得高分辨率的雄激素受体与其响应元件的复合物晶体结构，为前列腺癌的治疗提供药物靶点设计的理论基础.本研究构建了AR-DBD的原核表达载体， 在0.5 mmol/L IPTG，16 ℃条件下诱导24 h表达该蛋白.通过与AR响应元件寡核酸的结合优化蛋白纯化条件，使AR-DBD蛋白易于纯化.最终，蛋白核酸复合物晶体呈现出边缘清晰，短棒状的立体结构，本研究制备的蛋白核酸复合物晶体可直接用于X射线衍射分析.

Abstract:

Obtained a highresolution complex crystal structure of the androgen receptor (AR) and its response element (ARE) could provide a theoretical basis for drug target design for the treatment of prostate cancer. In this study, a prokaryotic expression vector of AR-DBD was constructed, and the protein was inducted to expression at 16 ℃ with 0.5 mmol/L IPTG for 24 h. In order to purify AR-DBD protein easily, the protein purification conditions were optimized by binding to ARE oligonucleic acids. Finally, we obtained the proteinnucleic acid complex crystal that exhibited short rodlike threedimensional structures with clear crystal edges. The proteinnucleotide complex crystals prepared in this study can be used for Xray diffraction analysis directly.