GPC3-Unc5 receptor complex structure and role in cell migration

Onno Akkermans; Céline Delloye-Bourgeois; Claudia Peregrina; Maria Carrasquero-Ordaz; Maria Kokolaki; Miguel Berbeira-Santana; Matthieu Chavent; Florie Reynaud; Ritu Raj; Jon Agirre; Metin Aksu; Eleanor S White; Edward Lowe; Dounia Ben Amar; Sofia Zaballa; Jiandong Huo; Irene Pakos; Patrick T N McCubbin; Davide Comoletti; Raymond J Owens; Carol V Robinson; Valérie Castellani; Daniel Del Toro; Elena Seiradake

doi:10.1016/j.cell.2022.09.025

GPC3-Unc5 receptor complex structure and role in cell migration

Cell. 2022 Oct 13;185(21):3931-3949.e26. doi: 10.1016/j.cell.2022.09.025.

Authors

Onno Akkermans¹, Céline Delloye-Bourgeois², Claudia Peregrina³, Maria Carrasquero-Ordaz⁴, Maria Kokolaki⁴, Miguel Berbeira-Santana⁴, Matthieu Chavent⁵, Florie Reynaud², Ritu Raj⁶, Jon Agirre⁷, Metin Aksu¹, Eleanor S White⁴, Edward Lowe¹, Dounia Ben Amar², Sofia Zaballa³, Jiandong Huo⁸, Irene Pakos⁹, Patrick T N McCubbin⁴, Davide Comoletti¹⁰, Raymond J Owens⁸, Carol V Robinson¹¹, Valérie Castellani¹², Daniel Del Toro¹³, Elena Seiradake¹⁴

Affiliations

¹ Department of Biochemistry, University of Oxford, Oxford, UK.
² MeLis, University of Lyon, Université Claude Bernard Lyon 1, CNRS UMR 5284, INSERM U1314, Institut NeuroMyoGène, 8 avenue Rockefeller 69008 Lyon, Lyon, France.
³ Department of Biological Sciences, Institute of Neurosciences, IDIBAPS, CIBERNED, University of Barcelona, Barcelona, Spain.
⁴ Department of Biochemistry, University of Oxford, Oxford, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK.
⁵ Institut de Pharmacologie et Biologie Structurale, Université de Toulouse, Toulouse, France.
⁶ Department of Chemistry, University of Oxford, Oxford, UK.
⁷ York Structural Biology Laboratory, Department of Chemistry, University of York, York, UK.
⁸ Structural Biology, The Rosalind Franklin Institute, Harwell Science Campus, Didcot, UK; Division of Structural Biology, University of Oxford, Oxford, UK.
⁹ Child Health Institute of New Jersey, New Brunswick, NJ 08901, USA.
¹⁰ Child Health Institute of New Jersey, New Brunswick, NJ 08901, USA; School of Biological Sciences, Victoria University of Wellington, Wellington, New Zealand.
¹¹ Department of Chemistry, University of Oxford, Oxford, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK.
¹² MeLis, University of Lyon, Université Claude Bernard Lyon 1, CNRS UMR 5284, INSERM U1314, Institut NeuroMyoGène, 8 avenue Rockefeller 69008 Lyon, Lyon, France. Electronic address: valerie.castellani@univ-lyon1.fr.
¹³ Department of Biological Sciences, Institute of Neurosciences, IDIBAPS, CIBERNED, University of Barcelona, Barcelona, Spain. Electronic address: danieldeltoro@ub.edu.
¹⁴ Department of Biochemistry, University of Oxford, Oxford, UK; Kavli Institute for Nanoscience Discovery, University of Oxford, Oxford, UK. Electronic address: elena.seiradake@bioch.ox.ac.uk.

Abstract

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

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Movement*
Glypicans / chemistry*
Glypicans / metabolism
Humans
Mice
Mutant Proteins
Netrin Receptors / chemistry*
Netrin Receptors / metabolism
Receptors, Cell Surface / metabolism
Single-Domain Antibodies
Thrombospondins

Substances

GPC3 protein, human
GPC3 protein, mouse
Glypicans
Mutant Proteins
Netrin Receptors
Receptors, Cell Surface
Single-Domain Antibodies
Thrombospondins
Unc5A protein, mouse
Unc5b protein, mouse

Grants and funding

202827/Z/16/Z/WT_/Wellcome Trust/United Kingdom