高迁移率蛋白B1免疫调控机制研究  

Study on the immunological regulation role of high mobility group box 1 protein

作　　者：赵擎[1] 姚咏明[2] 丁丽华[3] 叶棋浓[3] 

机构地区：[1]解放军总医院南楼消化内科,北京100853 [2]解放军总医院第304临床部烧伤研究所 [3]解放军军事医学科学院

出　　处：《中国医师杂志》2014年第1期6-9,共4页Journal of Chinese Physician

基　　金：国家自然科学基金（30801187）

摘　　要：目的 初步探讨高迁移率蛋白（HMG）B1与活化T细胞核因子（NFAT）2的结合区域.方法 首先构建HMGB1真核表达质粒,然后确定HMGB1分段位置,分别构建HMGB1的A box区域与B box区域真核表达质粒,以及NFAT2的真核表达质粒.通过蛋白纯化及体外翻译技术得到纯化蛋白,最后进行GST沉降实验,观察HMGB1区域蛋白与NFAT2蛋白的结合情况.结果 在谷胱甘肽转移酶-沉降实验中,可检测到HMGB1 B box融合蛋白与NFAT2结合的阳性条带,不能检测到HMGB1 A box融合蛋白与NFAT2结合的阳性条带.结论 HMGB1可通过B段区域与NFAT2发生相互作用.Objective To identify the specific binding-domain between high mobility group box-1 （HMGB1） protein and nu- clear factor of activated T cells-2 （NFAT2）. Methods Eukaryotic expression plasmids of HMGB1 A box （ 1-97 amino acid） , HG- MB1 B box （88 -166 amino acid）, and NFAT2 were constructed. Fragments of HMGB1 DNA were cloned into pGEX-KG vector. HMGB1 A box and B box proteins were prepared through protein expression and purification technique. Whole NFAT2 protein was pre- pared by reticulocyte transcription and translation system in vitro. The combination between HMGB1 A box protein, B box protein, and NFAT2 protein was detected with glutathione transferase （GST） -pull down assay. Results Interaction between HMGB1 B box protein and NFAT2 protein was confirmed, but not including the interaction between HMGB1 A box protein and NFAT2 protein. Conclusions HMGB1 could possibly bind to NFAT2 through B box domain.

关 键 词：高迁移率族蛋白质类 免疫学 转录因子 遗传载体 质粒 蛋白质类 分离和提纯 

分 类 号：R392[医药卫生—免疫学]