Soluble expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 in Escherichia coli

Jia Wei, Xiao Dan Cao, Sheng Min Zhou, Chao Chen, Hai Jun Yu, Yao Zhou, Ping Wang

Research output: Contribution to journalArticle

1 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1227-1233
Number of pages7
JournalJournal of Microbiology and Biotechnology
Volume25
Issue number8
DOIs
StatePublished - Apr 23 2015

Keywords

  • Anticancer drug screening
  • Catalytic domain
  • E. coli expression system
  • Inhibitors
  • Soluble expression
  • Vascular endothelial growth factor receptor 2

Fingerprint Dive into the research topics of 'Soluble expression and purification of the catalytic domain of human vascular endothelial growth factor receptor 2 in Escherichia coli'. Together they form a unique fingerprint.

  • Cite this