Abstract
Co-receptors add complexity to cell-cell signaling systems. The secreted semaphorin 3s (Sema3s) require a co-receptor, neuropilin (Nrp), to signal through plexin As (PlxnAs) in functions ranging from axon guidance to bone homeostasis, but the role of the co-receptor is obscure. Here we present the low-resolution crystal structure of a mouse semaphorin–plexin–Nrp complex alongside unliganded component structures. Dimeric semaphorin, two copies of plexin and two copies of Nrp are arranged as a dimer of heterotrimers. In each heterotrimer subcomplex, semaphorin contacts plexin, similar to in co-receptor–independent signaling complexes. The Nrp1s cross brace the assembly, bridging between sema domains of the Sema3A and PlxnA2 subunits from the two heterotrimers. Biophysical and cellular analyses confirm that this Nrp binding mode stabilizes a canonical, but weakened, Sema3–PlxnA interaction, adding co-receptor control over the mechanism by which receptor dimerization and/or oligomerization triggers signaling.
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Acknowledgements
We thank the staff of Diamond beamlines I02, I03 and I04-1 for assistance with diffraction data collection, Y. Zhao and W. Lu for help with protein expression, T.S. Walter for help with crystallization, G. Sutton for help with multiangle light scattering (MALS) experiments and A.R. Aricescu and D.I. Stuart for critical reading of the manuscript. This work was funded by Cancer Research UK (A10976) and the Medical Research Council (G9900061) to E.Y.J. B.J.C.J. is funded by the Human Frontier Science Program, G.A.W. and M.Z.C. are funded by the Medical Research Council and the John Fell Fund, C.S. is funded by the Wellcome Trust and E.Y.J. is funded by Cancer Research UK (A5261).
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B.J.C.J., T.M., C.S. and E.Y.J. designed the project. B.J.C.J. and E.Y.J. wrote the manuscript with input from all authors. B.J.C.J. produced the constructs for crystallization and performed MALS, crystallization, diffraction data collection, structure solution and refinement. T.M. and B.J.C.J. cloned and produced constructs for and performed SPR experiments. T.M. did the western blot analysis. T.M. produced proteins for growth cone–collapse assays, which G.A.W. performed under the supervision of M.Z.C.
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Janssen, B., Malinauskas, T., Weir, G. et al. Neuropilins lock secreted semaphorins onto plexins in a ternary signaling complex. Nat Struct Mol Biol 19, 1293–1299 (2012). https://doi.org/10.1038/nsmb.2416
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DOI: https://doi.org/10.1038/nsmb.2416
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