Abstract
Establishment of correct synaptic connections is a crucial step during neural circuitry formation. The Teneurin family of neuronal transmembrane proteins promotes cell-cell adhesion via homophilic and heterophilic interactions, and is required for synaptic partner matching in the visual and hippocampal systems in vertebrates. It remains unclear how individual Teneurins form macromolecular cis- and trans-synaptic protein complexes. Here, we present a 2.7 Å cryo-EM structure of the dimeric ectodomain of human Teneurin4. The structure reveals a compact conformation of the dimer, stabilized by interactions mediated by the C-rich, YD-shell, and ABD domains. A 1.5 Å crystal structure of the C-rich domain shows three conserved calcium binding sites, and thermal unfolding assays and SAXS-based rigid-body modeling demonstrate that the compactness and stability of Teneurin4 dimers are calcium-dependent. Teneurin4 dimers form a more extended conformation in conditions that lack calcium. Cellular assays reveal that the compact cis-dimer is compatible with homomeric trans-interactions. Together, these findings support a role for teneurins as a scaffold for macromolecular complex assembly and the establishment of cis- and trans-synaptic interactions to construct functional neuronal circuits.
| Original language | English |
|---|---|
| Article number | e107505 |
| Number of pages | 18 |
| Journal | EMBO Journal |
| Volume | 41 |
| Issue number | 9 |
| Early online date | 31 Jan 2022 |
| DOIs | |
| Publication status | Published - 2 May 2022 |
Bibliographical note
Funding Information:High-resolution EM data were collected at The Netherlands Centre for Electron Nanoscopy (NeCEN) with assistance from?Rebecca Dillard. X-ray measurements were performed on beamline ID23-1 at the European Synchrotron Radiation Facility (ESRF), Grenoble, France. We are grateful to David Flot at the ESRF for providing assistance in using beamline ID23-1. Scattering data were collected at Diamond Light Source (proposal mx19800) beamline B21 with assistance from Nathan Cowieson. K562 cells were a kind gift from Dr. Bas van Steensel at Netherlands Cancer Institute. We would like to thank Dr. Pouyan Boukany at Technical University of Delft for the electroporation device. B.J.C.J. is supported by a European Research Council starting grant (677500) and an NWO grant (OCENW.KLEIN.026). D.H.M. is supported by an NWO Veni grant (722.016.004) and a ZonMw Enabling Technology Hotel grant (435004008). The authors declare that they have no conflict of interest.
Funding Information:
High‐resolution EM data were collected at The Netherlands Centre for Electron Nanoscopy (NeCEN) with assistance from Rebecca Dillard. X‐ray measurements were performed on beamline ID23‐1 at the European Synchrotron Radiation Facility (ESRF), Grenoble, France. We are grateful to David Flot at the ESRF for providing assistance in using beamline ID23‐1. Scattering data were collected at Diamond Light Source (proposal mx19800) beamline B21 with assistance from Nathan Cowieson. K562 cells were a kind gift from Dr. Bas van Steensel at Netherlands Cancer Institute. We would like to thank Dr. Pouyan Boukany at Technical University of Delft for the electroporation device. B.J.C.J. is supported by a European Research Council starting grant (677500) and an NWO grant (OCENW.KLEIN.026). D.H.M. is supported by an NWO Veni grant (722.016.004) and a ZonMw Enabling Technology Hotel grant (435004008).
Publisher Copyright:
© 2022 The Authors. Published under the terms of the CC BY 4.0 license.
Keywords
- Teneurins
- neuroscience
- structural biology
- synaptic cell adhesion
- transmembrane proteins