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Updated: Jul 28, 2025

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APEX2-Mediated Proximity Labeling of Wnt Receptor Interactors Upon Pathway Activation

Ellen Youngsoo Rim1,2, Roeland Nusse1,2

  • 1Department of Developmental Biology, Stanford University School of Medicine.

Micropublication Biology
|June 1, 2023
PubMed

Insights

Researchers identified new proteins interacting with the Wnt signaling pathway receptor Frizzled 7 (Fzd7) upon Wnt stimulation using proximity-dependent biotinylation. These findings shed light on early Wnt signal transduction events.

Area of Science:

  • Molecular Biology
  • Cell Signaling
  • Developmental Biology

Background:

  • The Wnt signaling pathway is crucial for metazoan development, tissue homeostasis, and regeneration.
  • Understanding the molecular mechanisms immediately after Wnt-receptor interaction is essential for deciphering Wnt signal transduction.

Purpose of the Study:

  • To identify novel binding partners of the Wnt receptor Frizzled 7 (Fzd7) upon pathway activation.
  • To investigate the early molecular events in Wnt signal transduction.

Main Methods:

  • Frizzled 7 (Fzd7) was tagged with APEX2, an enzyme enabling proximity-dependent biotinylation.
  • APEX2-tagged Fzd7 localization and signaling activity were confirmed.
  • Biotinylation of proximal interactors was performed with and without Wnt3a stimulation, followed by mass spectrometry analysis.

Main Results:

  • Mass spectrometry identified known Wnt pathway proteins among the biotinylated interactors.
  • Proteins involved in actin cytoskeleton regulation, vesicle trafficking, and phospholipid modification were significantly enriched upon Wnt stimulation.
  • Several novel proteins with previously unestablished roles in Wnt signaling were identified in the Fzd7 interactome.

Conclusions:

  • The study successfully identified proximal interactors of Fzd7 in a Wnt-dependent manner.
  • The findings highlight the involvement of cytoskeleton regulation, vesicle trafficking, and phospholipid modification in early Wnt signaling.
  • Further investigation of newly identified proteins is warranted to elucidate their roles in the Wnt pathway.

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