一种高亲和结合Ⅰ型胶原蛋白的GST融合蛋白表达分析  

A GST fused protein expression with high affinity for type Ⅰ collagen

作　　者：董育红[1] 李遂焰[1] 陈渝萍[2,3] DONG Yu-hong;LI Sui-yan;CHEN Yu-ping(School of Life Science and Engineering,Southwest Jiaotong University,Chengdu 610031;School of Materials Science and Engineering,Southwest Jiaotong University,Chengdu 610031;School of Pharmaceutis and Life Sciences,University of South China,Hengyang 421001,China)

机构地区：[1]西南交通大学生命科学与工程学院,成都610031 [2]西南交通大学材料科学与工程学院,成都610031 [3]南华大学药学与生命科学学院,衡阳421001

出　　处：《生物学杂志》2019年第5期21-24,共4页Journal of Biology

基　　金：国家自然科学基金(81371675)

摘　　要：鉴于纤维化疾病病人组织中I型胶原的异常沉积和人胎盘生长因子2氨基酸123~144肽段(PlGF-2 123-144 )对I型胶原蛋白出众的高亲性,尝试制备PlGF-2 123-144 短肽用于改善抗纤维化药物的组织靶向特异性。首先对该短肽的编码DNA序列进行修改和扩增,构建了GST- PlGF-2 123-144 融合蛋白的原核表达质粒;进而使用 E.coli BL21诱导表达该融合蛋白,并以谷胱甘肽琼脂糖纯化后透析;最后采用ELISA检测确定,经原核表达和纯化所获得的GST- PlGF-2 123-14 融合蛋白具备对I型胶原蛋白的高亲性。为研发特异性靶向纤维化组织的抗纤维化药物打下了可靠基础。Given that type Ⅰ collagens dominate among ECMs aberrantly accumulated in fibrotic tissues, we consider to adopt the excellent affinity of the peptide of human PlGF-2 (aa. 123-144) for type Ⅰ collagens to improve tissue targeting specificity of antifibrotic agents. We firstly modified the DNA coding sequence of peptide PlGF-2 123-144 and constructed GST- PlGF-2 123-144 prokaryotic expression plasmid. GST-PlGF-2 123-144 fused proteins were then expressed in E. coli BL21, affinity purified with glutathione sepharose 4B matrix and PBS dialyzed. High affinity for type Ⅰ collagen of these prepared GST- PlGF-2 123-144 fused proteins was finally confirmed with ELISA assay. Our work contributes to the development of anti-fibrotic drugs that specifically target fibrotic tissue.

关 键 词：Ⅰ型胶原蛋白 PlGF-2 123-144 GST融合蛋白 原核表达 

分 类 号：Q78[生物学—分子生物学]