文献类型：期刊文献

英文题名：Soluble Expression and Purification of the Catalytic Domain of Human Vascular Endothelial Growth Factor Receptor 2 in Escherichia coli

作者：Wei, Jia[1];Cao, Xiaodan[1];Zhou, Shengmin[1];Chen, Chao[1];Yu, Haijun[1];Zhou, Yao[1];Wang, Ping[1]

机构：[1]E China Univ Sci & Technol, Shanghai Inst Biomfg Technol, Sch Biotechnol, State Key Lab Bioreactor Engn,Biomed Nanotechnol, Shanghai 200237, Peoples R China

年份：2015

卷号：25

期号：8

起止页码：1227

外文期刊名：JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000363180100005)】；

基金：This work was supported by the China Postdoctoral Science Foundation Grant (2013M540341), Key grant cultivating interdisciplinary studies of Chinese Education Ministry (WF1113014), National Natural Science Foundation of China (21303050 and 31471659), Pujiang Talent Program of Shanghai Municipality (13PJD012), and National "Thousand Talents Program" of China.

语种：英文

外文关键词：Vascular endothelial growth factor receptor 2; catalytic domain; soluble expression; E. coli expression system; inhibitors; anticancer drug screening

摘要：Vascular endothelial growth factor (VEGF) plays a key role in angiogenesis through binding to its specific receptors, which mainly occurs to VEGF receptor 2 (VEGFR-2), a kinase insert domain-containing receptor. Therefore, the disruption of VEGFR-2 signaling provides a promising therapeutic approach for the treatment of cancer by inhibiting abnormal or tumor-induced angiogenesis. To explore this potential, we expressed the catalytic domain of VEGFR2 (VEGFR-2-CD) as a soluble active kinase in Escherichia coli. The recombinant protein was purified and the VEGFR-2-CD activity was investigated. The obtained VEGFR-2-CD showed autophosphorylation activity and phosphate transfer activity comparable to the commercial enzyme. Furthermore, the IC50 value of known VEGFR-2 inhibitor was determined using the purified VEGFR-2-CD. These results indicated a possibility for functional and economical VEGFR-2-CD expression in E. coli to use for inhibitor screening.