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Quantitative Profiling of WNT-3A Binding to All Human Frizzled Paralogues in HEK293 Cells by NanoBiT/BRET Assessments

Kozielewicz, Paweł; Shekhani, Rawan; Moser, Stefanie; Bowin, Carl-Fredrik; Wesslowski, Janine; Davidson, Gary; Schulte, Gunnar


The WNT signaling system governs critical processes during embryonic development and tissue homeostasis, and its dysfunction can lead to cancer. Details concerning selectivity and differences in relative binding affinities of 19 mammalian WNTs to the cysteine-rich domain (CRD) of their receptors—the ten mammalian Frizzleds (FZDs)—remain unclear. Here, we used eGFP-tagged mouse WNT-3A for a systematic analysis of WNT interaction with every human FZD paralogue in HEK293A cells. Employing HiBiT-tagged full-length FZDs, we studied eGFP-WNT-3A binding kinetics, saturation binding, and competition binding with commercially available WNTs in live HEK293A cells using a NanoBiT/BRET-based assay. Further, we generated receptor chimeras to dissect the contribution of the transmembrane core to WNT-CRD binding. Our data pinpoint distinct WNT-FZD selectivity and shed light on the complex WNT-FZD binding mechanism. The methodological development described herein reveals yet unappreciated details of the complexity of WNT signaling and WNT-FZD interactions, providing further details with respect to WNT-FZD selectivity.

Verlagsausgabe §
DOI: 10.5445/IR/1000133876
Veröffentlicht am 14.06.2021
DOI: 10.1021/acsptsci.1c00084
Zitationen: 14
Zitationen: 17
Cover der Publikation
Zugehörige Institution(en) am KIT Institut für Biologische und Chemische Systeme (IBCS)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2021
Sprache Englisch
Identifikator ISSN: 2575-9108
KITopen-ID: 1000133876
HGF-Programm 43.33.11 (POF IV, LK 01) Adaptive and Bioinstructive Materials Systems
Erschienen in ACS Pharmacology and Translational Science
Verlag American Chemical Society (ACS)
Band 4
Heft 3
Seiten 1235–1245
Nachgewiesen in Scopus
Globale Ziele für nachhaltige Entwicklung Ziel 3 – Gesundheit und Wohlergehen
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