Quantitative Evaluation of VEGF Binding to VEGFR2 in Real Time in Living Cells
7 January 2019

Vascular endothelial growth factors (VEGF) are proteins that act as dimers to mediate angiogenesis by binding to their cellular receptors (VEGFR). Upon ligand binding VEGFRs dimerize and initiate a raft of signaling cascades via their intracellular tyrosine kinase domains. VEGFs and their receptors play a role in tumor vascularization and are major targets of anti-cancer therapies. In order to characterize affinities of VEGF isoforms binding to VEGFR2, assays are required that quantitatively and kinetically report on these interactions. A novel labeling technique stoichiometrically labels the VEGF family member VEGF165a with the fluorophore TMR. The modified ligand was used in conjunction with NanoLuciferase-coupled VEGFR2 (NLuc VEGFR2) in a cellular NanoBRET assay.