人CD96第一个IgV结构域的表达、结晶及晶体学分析

发布时间：2018-05-28 15:23

本文选题：CD + 第一IgV结构域　；参考：《生物工程学报》2013年05期

【摘要】：CD96(Tactile)是一个主要表达在活化的T细胞、NK细胞上的免疫受体分子。它属于免疫球蛋白超家族,胞外区有3个免疫球蛋白结构域(V1,V2/C和C)。最近的研究表明,CD96的第一个IgV结构域(CD96V1)在细胞粘附和NK细胞介导的杀伤过程中起着重要的作用。文中构建了人类CD96第一个IgV样结构域(hCD96V1)的表达载体,并将它在大肠杆菌BL21(DE3)菌株中以包涵体形式表达。通过包涵体体外复性,得到了CD96可溶性蛋白。分析超速离心结果证明CD96可溶蛋白在体外以二聚体形式存在。采用座滴气相扩散法获得了衍射质量较好的CD96蛋白晶体,衍射分辨率为1.9,晶体属于空间群P21,晶胞参数为a=35.1,B=69.5,C=49.6,α=γ=90°,β=105.4°。CD96晶体及衍射数据的获得为后续的结构和功能研究奠定了基础。
[Abstract]:CD96 Tactile is an immunoreactive receptor molecule mainly expressed on activated T cell / NK cells. It belongs to the immunoglobulin superfamily. There are three immunoglobulin domains V _ 1 V _ (2 / C) and C _ (2 / C) in the extracellular domain. Recent studies have shown that the first IgV domain of CD96, CD96V1, plays an important role in cell adhesion and NK cell-mediated killing. The expression vector of the first IgV like domain hCD96V1 of human CD96 was constructed and expressed as inclusion body in E. coli BL21DDE3 strain. CD96 soluble protein was obtained by in vitro renaturation of inclusion bodies. The results of ultracentrifugation showed that CD96 soluble protein existed in the form of dimer in vitro. The crystal of CD96 protein with good diffraction quality was obtained by gas phase diffusion method. The diffraction resolution is 1.9, the crystal belongs to space group P21, the crystal cell parameters are: a ~ (35.1) ~ (35. 1) B ~ (2 +) 69.5C ~ (1), 伪 = 纬 ~ (2 +) 90 掳, 尾 ~ (10) 4 掳路CD ~ (96), and the obtained diffraction data lay a foundation for the further study of structure and function.
【作者单位】：安徽农业大学生命科学学院;中国科学院微生物研究所中国科学院病原微生物与免疫学重点实验室;
【基金】：国家自然科学基金(No.31030030)资助~~
【分类号】：R392

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