Novel Bioluminescent Binding Assays for Ligand–Receptor Interaction Studies of the Fibroblast Growth Factor Family
Fig 4
Binding of FGF2-Luc with the endogenous FGF receptor.
Untransfected HEK293T cells were used as the receptor source. (A) Saturation binding of FGF2-Luc with the endogenous FGF receptor. Nonspecific binding data were obtained by competition with 250 nM of 6×His-FGF2. The measured bioluminescence data were expressed as mean ± SE (n = 3). The total binding data were fitted to Y = BmaxX/(Kd + X) + NsX, specific binding data to Y = BmaxX/(Kd + X), and nonspecific binding data to a linear curve. Inner panel, Scatchard plot of the specific binding data. (B) Competition binding of wild-type or mutant FGF2s with the endogenous FGF receptor using FGF2-Luc as a tracer. The measured bioluminescence data were expressed as mean ± SE (n = 3) and fitted with sigmoidal curves using the SigmaPlot10.0 software.