<br/>Fibroblast growth factors (FGF) are heparin binding proteins that signal through their fibroblast growth factor receptors (FGFR) via interaction with immunoglobin-like domains. FGFs regulate cellular processes that leads to cell proliferation via angiogenesis, which is particularly important in wound healing. It is hypothesized that signaling involves FGF binding to heparin on the extracellular matrix together with binding to the FGFR leading to dimerization, activation of tyrosine kinase and subsequently autophosphorylation which activates signal transduction. Deregulation of FGF signaling has been found to be responsible for excessive cell proliferation found in many cancers. As a result, understanding the signaling of FGF has become an important target for cancer therapy. We are studying the interaction between FGF and FGFR by fluorescence resonance energy transfer (FRET). In FRET, one molecule is site-specifically labelled with a donor and the other with an acceptor using maleimide dyes, which attach to the proteins via cysteine residues incorporated into the protein at specific sites. When the proteins interact, the donor will transfer its energy to the acceptor, where the efficiency can be used to extract both structural and kinetic information about the interaction. Here we provide preliminary results regarding the design, mutation, expression and site-specific labeling of FGFR and FGF. Our preliminary results show successful mutants of FGF and FGFR where we use AlexFluor594 as the acceptor dye on FGF and AlexaFluo488 as the donor dye on FGFR. We discuss how FRET experiments are being used to provide further understanding on the binding mechanisms of FGF and FGFR.

Development of FRET assays for investigating the interaction between fibroblast growth factor (FGF) and its receptor (FGFR)

251st National Meeting (2016)

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