Super-complexes of adhesion GPCRs and neural guidance receptors.
Latrophilin 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.
Department of Biochemistry, Oxford University, Oxford OX1 3QU, UK.
Verity A
VA
Jackson
Department of Chemistry, University of Oxford, Oxford OX1 3QZ, UK.
Shahid
S
Mehmood
Department of Biochemistry, Oxford University, Oxford OX1 3QU, UK.
Matthieu
M
Chavent
Department of Biochemistry, Oxford University, Oxford OX1 3QU, UK.
Pietro
http://orcid.org/0000-0001-9280-9437
P
Roversi
Department of Biochemistry, Oxford University, Oxford OX1 3QU, UK.
Maria
M
Carrasquero
Max-Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Munich-Martinsried, Germany.
Daniel
D
Del Toro
Max-Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Munich-Martinsried, Germany.
Goenuel
G
Seyit-Bremer
Child Health Institute of New Jersey, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, New Jersey 08901, USA.|Department of Neuroscience and Cell Biology, Rutgers University, New Brunswick, New Jersey 08901, USA.|Department of Pediatrics, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, New Jersey 08901, USA.
Fanomezana M
FM
Ranaivoson
Child Health Institute of New Jersey, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, New Jersey 08901, USA.|Department of Neuroscience and Cell Biology, Rutgers University, New Brunswick, New Jersey 08901, USA.|Department of Pediatrics, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, New Jersey 08901, USA.
Davide
D
Comoletti
Department of Biochemistry, Oxford University, Oxford OX1 3QU, UK.
Mark S P
MS
Sansom
Department of Chemistry, University of Oxford, Oxford OX1 3QZ, UK.
Carol V
CV
Robinson
Max-Planck Institute of Neurobiology, Am Klopferspitz 18, 82152 Munich-Martinsried, Germany.|Munich Cluster for Systems Neurology (SyNergy), Feodor-Lynen-Straβe 17, 81377 Munich, Germany.
Rüdiger
R
Klein
Department of Biochemistry, Oxford University, Oxford OX1 3QU, UK.
Elena
E
Seiradake
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D018360:Crystallography, X-Ray; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry; D008562:Membrane Glycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018345:Mice, Knockout; D056004:Molecular Dynamics Simulation; D008969:Molecular Sequence Data; D046912:Multiprotein Complexes / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018000:Receptors, Peptide / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D020349:Surface Plasmon Resonance
D008562:Membrane Glycoproteins; D046912:Multiprotein Complexes; D011956:Receptors, Cell Surface; D043562:Receptors, G-Protein-Coupled; D018000:Receptors, Peptide; C514259:FLRT2 protein, mouse; C000592543:Adgrl3 protein, mouse; C572338:Unc5d protein, mouse