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[1]张兰君,靳林丹,任海霞,等.人源Pelota C端结构域的纯化、结晶及晶体结构分析[J].厦门大学学报(自然科学版),2017,56(01):64-71.[doi:10.6043/j.issn.0438-0479.201604036]
 ZHANG Lanjun,JIN Lindan,REN Haixia,et al.Purification,Crystallization and Crystallographic Analysis of the Pelota C-terminal Domain from Human[J].Journal of Xiamen University(Natural Science),2017,56(01):64-71.[doi:10.6043/j.issn.0438-0479.201604036]
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人源Pelota C端结构域的纯化、结晶及晶体结构分析(PDF)
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《厦门大学学报(自然科学版)》[ISSN:0438-0479/CN:35-1070/N]

卷:
56卷
期数:
2017年01期
页码:
64-71
栏目:
研究论文
出版日期:
2017-01-23

文章信息/Info

Title:
Purification,Crystallization and Crystallographic Analysis of the Pelota C-terminal Domain from Human
文章编号:
0438-0479(2017)01-0064-08
作者:
张兰君靳林丹任海霞林天伟*
厦门大学生命科学学院,福建 厦门 361102
Author(s):
ZHANG LanjunJIN LindanREN HaixiaLIN Tianwei*
School of Life Sciences,Xiamen University,Xiamen 361102,China
关键词:
Pelota C端结构域 人源 表达 纯化 结晶
Keywords:
Pelota C-terminal domain human expression purification crystallization
分类号:
Q 356.1
DOI:
10.6043/j.issn.0438-0479.201604036
文献标志码:
A
摘要:
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0.26 nm分辨率.蛋白晶体的空间群为P6522,晶胞常数a=7.882 nm,b=7.882 nm,c=19.746 nm.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.
Abstract:
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 filtration chromatography.Finally,the protein is over 97% in purity.Dynamic light scattering experiments showed that the purified protein 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 P6522,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.

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备注/Memo

备注/Memo:
收稿日期:2016-04-18 录用日期:2016-06-28
基金项目:高等学校学科创新引智计划(“111”计划)(B06016)
*通信作者:twlin@xmu.edu.cn
更新日期/Last Update: 1900-01-01