IRF6调控IFN-β的功能验证与结构研究Study on the interferon-β regulation and structure of IRF6
魏迪洋;闫利明;李巍然;贾绮林;
摘要(Abstract):
利用X射线衍射晶体学方法解析IRF6蛋白的三维结构,同时验证IRF6对干扰素-β的转录调控作用,为IRF6蛋白的结构生物学研究提供基础。研究利用高通量基因克隆、蛋白表达与纯化获得大量的IRF6 N端蛋白结构域可溶性表达。凝胶迁移试验(EMSA)结果显示,IRF6 N端结构域可与干扰素-β启动子核酸片段形成蛋白-核酸复合体,具有明确的相互作用。将孵育后的蛋白-核酸复合体进行悬滴气相扩散法晶体筛选与优化后获得了X射线衍射分辨率达2.68?的晶体,为进一步揭示IRF6蛋白三维结构提供了帮助。验证了IRF6具有结合干扰素-β启动子元件特定片段的N端结构域,可调控干扰素-β的转录与表达,并通过IRF6 N端结构域与干扰素-β启动子核酸片段共同孵育后获得高分辨率蛋白晶体,以解析IRF6蛋白三维结构。研究结果对IRF6相关疾病机制研究及干扰素相关免疫通路的调控机制有重要意义。
关键词(KeyWords): 干扰素调节因子6;干扰素-β;蛋白纯化;蛋白结晶
基金项目(Foundation): 国家自然科学基金青年项目(3170040488)
作者(Author): 魏迪洋;闫利明;李巍然;贾绮林;
Email:
DOI:
参考文献(References):
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