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

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 OwensCarol V Robinson, Valérie Castellani, Daniel Del Toro, Elena Seiradake

Chemistry

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Pages (from-to)	3931-3949.e26
Number of pages	46
Journal	Cell
Volume	185
Issue number	21
DOIs	https://doi.org/10.1016/j.cell.2022.09.025
Publication status	Published - 13 Oct 2022

Bibliographical note

Keywords

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

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10.1016/j.cell.2022.09.025Licence: CC BY

GPC3-Unc5 receptor complex structure and role in cell migration
© 2022 The Authors
Final published version, 16 MBLicence: CC BY

URF Enhancement Award - Years 1, 2 and 3
Agirre, J. (Principal investigator)
THE ROYAL SOCIETY
1/10/22 → 30/09/25
Project: Research project (funded) › Research

Cite this

Akkermans, O., Delloye-Bourgeois, C., Peregrina, C., Carrasquero-Ordaz, M., Kokolaki, M., Berbeira-Santana, M., Chavent, M., Reynaud, F., Raj, R., Agirre, J., Aksu, M., White, E. S., Lowe, E., Ben Amar, D., Zaballa, S., Huo, J., Pakos, I., McCubbin, P. T. N., Comoletti, D., ... Seiradake, E. (2022). GPC3-Unc5 receptor complex structure and role in cell migration. Cell, 185(21), 3931-3949.e26. https://doi.org/10.1016/j.cell.2022.09.025

@article{048ff411cb79402eb761ab995119e64a,

title = "GPC3-Unc5 receptor complex structure and role in cell migration",

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.",

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doi = "10.1016/j.cell.2022.09.025",

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Akkermans, O, Delloye-Bourgeois, C, Peregrina, C, Carrasquero-Ordaz, M, Kokolaki, M, Berbeira-Santana, M, Chavent, M, Reynaud, F, Raj, R, Agirre, J, Aksu, M, White, ES, Lowe, E, Ben Amar, D, Zaballa, S, Huo, J, Pakos, I, McCubbin, PTN, Comoletti, D, Owens, RJ, Robinson, CV, Castellani, V, Del Toro, D & Seiradake, E 2022, 'GPC3-Unc5 receptor complex structure and role in cell migration', Cell, vol. 185, no. 21, pp. 3931-3949.e26. https://doi.org/10.1016/j.cell.2022.09.025

TY - JOUR

T1 - GPC3-Unc5 receptor complex structure and role in cell migration

AU - Akkermans, Onno

AU - Delloye-Bourgeois, Céline

AU - Peregrina, Claudia

AU - Carrasquero-Ordaz, Maria

AU - Kokolaki, Maria

AU - Berbeira-Santana, Miguel

AU - Chavent, Matthieu

AU - Reynaud, Florie

AU - Raj, Ritu

AU - Agirre, Jon

AU - Aksu, Metin

AU - White, Eleanor S

AU - Lowe, Edward

AU - Ben Amar, Dounia

AU - Zaballa, Sofia

AU - Huo, Jiandong

AU - Pakos, Irene

AU - McCubbin, Patrick T N

AU - Comoletti, Davide

AU - Owens, Raymond J

AU - Robinson, Carol V

AU - Castellani, Valérie

AU - Del Toro, Daniel

AU - Seiradake, Elena

PY - 2022/10/13

Y1 - 2022/10/13

N2 - 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.

AB - 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.

KW - Animals

KW - Cell Movement

KW - Glypicans/chemistry

KW - Humans

KW - Mice

KW - Mutant Proteins

KW - Netrin Receptors/chemistry

KW - Receptors, Cell Surface/metabolism

KW - Single-Domain Antibodies

KW - Thrombospondins

U2 - 10.1016/j.cell.2022.09.025

DO - 10.1016/j.cell.2022.09.025

M3 - Article

C2 - 36240740

SN - 0092-8674

VL - 185

SP - 3931-3949.e26

JO - Cell

JF - Cell

IS - 21

ER -

GPC3-Unc5 receptor complex structure and role in cell migration

Abstract

Bibliographical note

Keywords

Access to Document

Projects

URF Enhancement Award - Years 1, 2 and 3

Cite this