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Expression and purification of human vascular‐endothelial‐growth‐factor‐receptor‐2 tyrosine kinase in Streptomyces for inhibitor screening
Author(s) -
Liu Chunping,
Guo Lianhong,
Yao Chen,
Zhang Ren,
Li Yuan
Publication year - 2008
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1042/ba20070112
Subject(s) - kinase insert domain receptor , angiogenesis , receptor tyrosine kinase , tyrosine kinase , vascular endothelial growth factor , biology , vascular endothelial growth factor a , biochemistry , microbiology and biotechnology , receptor , chemistry , cancer research , vegf receptors
VEGF (vascular endothelial growth factor) is a critical regulator in angiogenesis through binding to its specific receptors, including VEGFR‐2 (VEGF receptor 2), a kinase insert domain‐containing receptor, on the surface of endothelial cells. As angiogenesis has been shown to be essential for malignancy of tumours, VEGFR‐2 is a potential therapeutic target for the treatment of cancers. To explore this potential, VEGFR‐2‐CD (the protein tyrosine kinase catalytic domain of VEGFR‐2) was cloned and expressed in Streptomyces lividans TK24, a prokaryotic expression system. The recombinant protein was purified, and correlations between its activity and enzyme concentration, ATP concentration, substrate concentration and bivalent cations were characterized. An ELISA‐based screening system was then established and used to search for inhibitors acting on the tyrosine kinase part of VEGFR‐2. More than 600 compounds originating from a variety of microbes have been screened so far, and a number of them have been demonstrated to be potential inhibitors of VEGFR‐2 TK.