Super-complexes of adhesion GPCRs and neural guidance receptors.
journal contribution
posted on 2019-09-09, 09:20 authored by VA Jackson, S Mehmood, M Chavent, P Roversi, M Carrasquero, D Del Toro, G Seyit-Bremer, FM Ranaivoson, D Comoletti, MSP Sansom, CV Robinson, R Klein, E SeiradakeLatrophilin adhesion-GPCRs (Lphn1-3 or ADGRL1-3) and Unc5 cell guidance receptors (Unc5A-D) interact with FLRT proteins (FLRT1-3), thereby promoting cell adhesion and repulsion, respectively. How the three proteins interact and function simultaneously is poorly understood. We show that Unc5D interacts with FLRT2 in cis, controlling cell adhesion in response to externally presented Lphn3. The ectodomains of the three proteins bind cooperatively. Crystal structures of the ternary complex formed by the extracellular domains reveal that Lphn3 dimerizes when bound to FLRT2:Unc5, resulting in a stoichiometry of 1:1:2 (FLRT2:Unc5D:Lphn3). This 1:1:2 complex further dimerizes to form a larger 'super-complex' (2:2:4), using a previously undescribed binding motif in the Unc5D TSP1 domain. Molecular dynamics simulations, point-directed mutagenesis and mass spectrometry demonstrate the stability and molecular properties of these complexes. Our data exemplify how receptors increase their functional repertoire by forming different context-dependent higher-order complexes.
Funding
We thank M. Jones, K. Harlos, D. Staunton and E. Lowe for technical support, the Diamond Light Source and ESRF for beamtime (proposals mx8423, mx9306, mx1747), the staff of beamlines I04-1 and ID-29, C. Vonrhein and tutors at the 2015 Diamond-CCP4 course for advice on data processing and refinement. We thank E.Y. Jones for initial crystallographic facilities and N. Zitzmann for computer facilities. M.Ch. thanks C. Song for fruitful discussion. We thank J. Egea for a construct of FLRT2 with a FLAG tag sequence at the C terminus. This work was funded by the UK Medical Research Council (MR/L018039/1), the John Fell OUP Research Fund, the Max-Planck Society, and the Deutsche Forschungsgemeinschaft SFB 834 and EXC 115. D.dT. was funded by a Marie Curie IEF fellowship (ID 274541). V.A.J was supported by a Wellcome Trust DPhil studentship. D.C. was supported by National Institutes of Health grant MH092906 and grant 67038 from the Robert Wood Johnson Foundation to the Child Health Institute of New Jersey. Research in MSPS’s group is supported by the Wellcome Trust (WT092970MA). S.M. and C.V.R. acknowledge support from a Medical Research Council Program grant (98101). Molecular Dynamics simulations were performed using the ARCHER supercomputer via the EPSRC-funded HECBioSim consortium.
History
Citation
Nature Communications, 2016, 7, Article number: 11184Author affiliation
/Organisation/COLLEGE OF LIFE SCIENCES/Biological Sciences/Molecular & Cell BiologyVersion
- VoR (Version of Record)
Published in
Nature CommunicationsPublisher
Nature Research (part of Springer Nature)eissn
2041-1723Acceptance date
2016-02-26Copyright date
2016Available date
2019-09-09Publisher DOI
Publisher version
https://www.nature.com/articles/ncomms11184Notes
Accession codes: The RCSB PDB accession number for the tetrameric and octameric Lphn3:FLRT2:Unc5D complexes reported in this paper are 5ftu and 5ftt, respectively. Supplementary Information accompanies this paper at http://www.nature.com/naturecommunicationsLanguage
enAdministrator link
Usage metrics
Categories
No categories selectedKeywords
Amino Acid SequenceAnimalsCell AdhesionCrystallography, X-RayHEK293 CellsHeLa CellsHumansMass SpectrometryMembrane GlycoproteinsMice, KnockoutMolecular Dynamics SimulationMolecular Sequence DataMultiprotein ComplexesProtein BindingProtein MultimerizationProtein Structure, SecondaryProtein Structure, TertiaryReceptors, Cell SurfaceReceptors, G-Protein-CoupledReceptors, PeptideSequence Homology, Amino AcidSurface Plasmon Resonance
Licence
Exports
RefWorks
BibTeX
Ref. manager
Endnote
DataCite
NLM
DC