eGFP-tagged Wnt-3a enables functional analysis of Wnt trafficking and signaling and kinetic assessment of Wnt binding to full-length Frizzled
Janine Wesslowski, Paweł Kozielewicz, Xianxian Wang, Haijun Cui, Hannes Schihada, Dominique Kranz, Pradhipa Karuna M, Pavel A. Levkin, Julia Christina Gross, Michael Boutros, Gunnar Schulte, Gary Davidson
Abstract
embryos, stimulated low-density lipoprotein receptor-related protein 6 (LRP6) phosphorylation in cells, and associated with exosomes. Further, we used conditioned medium containing eGFP-Wnt-3a to visualize its binding to FZD and to quantify Wnt-FZD interactions in real time in live cells, utilizing a recently established NanoBRET-based ligand binding assay. In summary, the development of a biologically active, fluorescent Wnt-3a reported here opens up the technical possibilities to unravel the intricate biology of Wnt signaling and Wnt-receptor selectivity.