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http://dx.doi.org/10.4014/jmb.1503.03073

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

Wei, Jia (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Cao, Xiaodan (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Zhou, Shengmin (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Chen, Chao (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Yu, Haijun (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Zhou, Yao (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Wang, Ping (State Key Laboratory of Bioreactor Engineering, Biomedical Nanotechnology Center, Shanghai Institute of Biomanufacturing Technology, School of Biotechnology, East China University of Science and Technology)
Publication Information
Journal of Microbiology and Biotechnology / v.25, no.8, 2015 , pp. 1227-1233 More about this Journal
Abstract
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 tumorinduced angiogenesis. To explore this potential, we expressed the catalytic domain of VEGFR-2 (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.
Keywords
Vascular endothelial growth factor receptor 2; catalytic domain; soluble expression; E. coli expression system; inhibitors; anticancer drug screening;
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