Abstract
Fibroblast growth factor receptors (FGFRs) are a family of receptor tyrosine kinases containing three domains: an extracellular receptor domain, a single transmembrane helix, and an intracellular tyrosine kinase domain. FGFRs are activated by fibroblast growth factors (FGFs) as part of complex signal transduction cascades regulating angiogenesis, skeletal formation, cell differentiation, proliferation, cell survival, and cancer. We have developed the first recombinant expression system in E. coli to produce a construct of human FGFR2 containing its transmembrane and extracellular receptor domains. We demonstrate that the expressed construct is functional in binding heparin and dimerizing. Size exclusion chromatography demonstrates that the purified FGFR2 does not form a complex with FGF1 or adopts an inactive dimer conformation. Progress towards the successful recombinant production of intact FGFRs will facilitate further biochemical experiments and structure determination that will provide insight into how extracellular FGF binding activates intracellular kinase activity.
Highlights
As receptor tyrosine kinases (RTKs), Fibroblast growth factor receptors (FGFRs) have three primary domains: an extracellular domain (ECD), a single transmembrane helix (TM), and an intracellular tyrosine kinase domain (KD) (Fig. 1)
We show that the FGFR2 ECD + TM construct is functional in binding heparin and dimerizing
We considered the FGFR2 and FGFR3 ECD + TM constructs to be the most promising for larger scale expression studies because of their superior yield, and the partial recovery of soluble FGFR3 ECD + TM
Summary
As receptor tyrosine kinases (RTKs), Fibroblast growth factor receptors (FGFRs) have three primary domains: an extracellular domain (ECD), a single transmembrane helix (TM), and an intracellular tyrosine kinase domain (KD) (Fig. 1). These proteins are expressed primarily in endothelial, fibroblast, vascular smooth muscle, neuroectodermal, and mesenchymal cells. Within the subfamily are four types of FGFRs: FGFR1, FGFR2, FGFR3, and FGFR4, which share 55–72% sequence homology Due to their critical roles in cell and tissue development, mutations of FGFRs are known to lead to achondroplasia (poor cartilage growth) and developmental disorders that exhibit craniosynostosis (improper skull formation) (Turner & Grose, 2010). FGFR2 and FGFR3 have been implicated in cancers such as bladder
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