Cell
Volume 185, Issue 21, 13 October 2022, Pages 3931-3949.e26
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Article
GPC3-Unc5 receptor complex structure and role in cell migration

https://doi.org/10.1016/j.cell.2022.09.025Get rights and content
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Highlights

  • Uncoordinated-5 receptor D (Unc5D) and glypican-3 (GPC3) form an octameric complex

  • Anti-GPC3 nanobodies (Nanoglue, Nanobreak) enhance or weaken Unc5-GPC3 binding

  • Unc5/GPC3 signaling causes cell contact-repulsion in vitro

  • In vivo, Unc5/GPC3 modulate the migration of cortical neurons and neuroblastoma cells

Summary

Neural migration is a critical step during brain development that requires the interactions of cell-surface guidance receptors. Cancer cells often hijack these mechanisms to disseminate. Here, we reveal crystal structures of Uncoordinated-5 receptor D (Unc5D) in complex with morphogen receptor glypican-3 (GPC3), forming an octameric glycoprotein complex. In the complex, four Unc5D molecules pack into an antiparallel bundle, flanked by four GPC3 molecules. Central glycan-glycan interactions are formed by N-linked glycans emanating from GPC3 (N241 in human) and C-mannosylated tryptophans of the Unc5D thrombospondin-like domains. MD simulations, mass spectrometry and structure-based mutants validate the crystallographic data. Anti-GPC3 nanobodies enhance or weaken Unc5-GPC3 binding and, together with mutant proteins, show that Unc5/GPC3 guide migrating pyramidal neurons in the mouse cortex, and cancer cells in an embryonic xenograft neuroblastoma model. The results demonstrate a conserved structural mechanism of cell guidance, where finely balanced Unc5-GPC3 interactions regulate cell migration.

Keywords:

uncoordinated-5
Unc5
glypican-3
GPC3
crystallography
structural biology
cell guidance
cell migration
cortex development
neuroblastoma
stripe assay
nanobodies
surface plasmon resonance
UNC5A
UNC5B
UNC5C
UNC5D

Data and code availability

  • Crystallography data and models have been deposited at the PDB where they will be publicly available as of the date of publication. Accession numbers are listed in the key resources table. This paper also analyses existing, publicly available RNA sequencing data. Accession numbers are listed in the key resources table.

  • This paper does not report original code.

  • Any additional information required to reanalyze the data reported in this paper is available from the lead contact upon request.

Cited by (0)

12

These authors contributed equally

13

Lead contact