人源Pelota C端结构域的纯化、结晶及晶体结构分析

Purification,Crystallization and Crystallographic Analysis of the Pelota C-terminal Domain from Human

Pelota在进化上是非常保守的RNA结合蛋白,人源Pelota mRNA分布于几乎所有的组织并作为一个多功能的蛋白参与多种生物途径.为解析人源Pelota C端结构域(C-terminal domain,CTD)的晶体结构,首先在大肠杆菌(Escherichia coli)中表达,并采用亲和层析、凝胶过滤柱层析的方法,获得了纯度大于97%的蛋白.动态光散射实验表明纯化的蛋白有较高的均一性.在筛选了1 852个结晶条件后,优化的人源Pelota CTD蛋白晶体能衍射X射线至0.26nm分辨率.蛋白晶体的空间群为P6522,晶胞常数a=7.882nm,b=7.882nm,c=19.746nm.上述结果为进一步研究Pelota的功能及其与下游蛋白的相互作用奠定了结构基础.

Pelota, an evolutionarily conservative RNA binding protein,is distributed in almost all tissues and involved in a variety of cell biological regulation as a multifunctional protein.In order to determine the crystal structure of the human Pelota C domain （C- terminal domain,CTD） ,we chose Escherichia coli to express the protien and purified the protien by affinity chromatography,gel fil- tration chromatography.Finally, the protein is over 97 % in purity.Dynamic light scattering experiments showed that the purified pro- tein had high homogeneity.After screening the 1 852 crystallization conditions, the optimized crystal of the human Pelota C domain can be diffracted to 0.26 nm resolution, The space group of the crystal is P6s 22 ,and the unit cell constant is a =7. 882 nm,b = 7. 882 nm,c=19. 746 nm.We determine the crystal structure of human Pelota CTD in this study,which lays a foundation for further study on the function of Pelota protein and its interaction with the downstream proteins.