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热休克因子结合蛋白（HSBP1）的表达、纯化、结晶与初步晶体学研究

ISSN号：1000-3282
期刊名称：《生物化学与生物物理进展》
时间：0
分类：Q71[生物学—分子生物学]
作者机构：[1]清华大学结构生物学实验室,北京100084, [2]中国科学院生物物理研究所,生物大分子国家重点实验室,北京100101
相关基金：supported by grants from The Ministry of Science and Technology of China （2006DFB32420）; The National Natural Science Foundation of China （30221003 30670494）; The “Sino-European Project on SARS Diagnostics and Antivirals” （SEPSDA） of The European Commission （003831）

作者：张红彦[1], 刘学琦[2], 孙玉娜[2], 郝宁[2], 李雪梅[2], 娄智勇[1]

关键词：热休克因子结合蛋白(HSBP1), 纯化, 结晶, heat shock factor binding protein 1 （HSBP1）, purification, crystallization

中文摘要：

热休克反应（heat shock response,HSR）是细胞在缺氧、病毒感染等应激因素刺激下,为适应微环境的改变而发生的一种自身保护性反应,以暂时性下调细胞正常代谢和选择性上调热休克蛋白（heat shock protein,HSP）的表达为特点.HSR是通过热休克转录因子（heat shock transcription factor,HSF）与相应的启动子结合,启动转录过程,进而促使HSP的表达来实现,其中HSF1是最有代表性、研究最多的一种HSF.HSF1在无活性的单体及有活性的三聚体之间的转变和平衡是转录调控的关键.热休克因子结合蛋白（heat shock factor binding protein,HSBP1）是一个含有两个伸展的疏水重复区,与HSF1的三聚体区域相互作用,以其伴侣形式实现对HSF1的DNA结合活性有负调节作用,通过抑制HSF1与DNA结合活性,从而抑制HSF1的转录活性.为了深入研究HSBP1行使功能的结构基础,对HSBP1蛋白成功地进行了克隆表达和结晶,该晶体属于R3空间群,其晶胞参数为a=b=35.2,c=233.3.

英文摘要：

Heat shock response rises when the organisms in the environmental stimulate. During heat shock, a lot of proteins which is so called heat shock protein that do not express in normal state largely express. These proteins′ expression is regulated by a family of transcription factor. These proteins were called heat shock transcriptional factors （HSFs）. Among these HSFs, HSF1 is the most important protein. And HSFs are also regulated by other proteins, such as heat shock factor binding protein1 （HSBP1）. HSBP1 interact with the trimer formation of HSF1 to convert the HSF1 from its active trimer state to inert monomer state. In order to do the further functional investigation, the gene of HSBP1 was cloned by PCR using the virus cDNAs as templates and expressed in Escherichia coli BL21 （DE3）. Furthermore, the expressed HSBP1 protein was purified and crystallized. The distinct crystal form was obtained by the hanging-drop vapor-diffusion process to carry on screening and the optimization to the crystallization condition. The crystals belong to R3 space group with the cell parameter a = b = 35.2, c = 233.3.