Distribution, assembly and mechanism of GluN1/GluN3A excitatory glycine receptors

Xu, L.; de battista, M.; Yao, K.; Schwenk, J.; Nehme, L.; pizzamiglio, l.; Cerasale, A.; Fakler, B.; Stroebel, D.; Zhu, S.; Paoletti, P.

doi:10.64898/2026.03.29.715060

bioRxiv · 10.64898/2026.03.29.715060

Distribution, assembly and mechanism of GluN1/GluN3A excitatory glycine receptors

Xu, L.·de battista, M.·Yao, K.·Schwenk, J.·Nehme, L.·pizzamiglio, l.·Cerasale, A.·Fakler, B.·Stroebel, D.·Zhu, S.·Paoletti, P.

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Abstract

NMDA receptors play key roles in brain development, plasticity and diseases. While glutamate and glycine co-gated GluN2-containing NMDARs have been extensively characterized, little is known regarding GluN3A-containing NMDARs that form receptors gated by glycine only. Here, combining native purification, mass spectrometry, cryo-EM and electrophysiology, we provide key insights on the molecular logic of GluN3A-NMDARs. We demonstrate that native GluN3A receptors account for a sizeable fraction of total NMDARs, are enriched at extrasynaptic compartments in the adult brain, and assemble specifically as diheteromeric GluN1/GluN3A excitatory glycine receptors (eGlyRs) rather than as triheteromeric GluN1/GluN2/GluN3A receptors. The architecture of eGlyRs reveal splayed and loosely packed extracellular domains, strikingly different from conventional GluN1/GluN2 receptors. Through back-and- forth structural and functional validations, we demonstrate how the combined effects of a weak ligand-binding domain (LBD) dimer interface and high intrinsic mobility of the N-terminal domains (NTDs) shape the atypical gating of eGlyRs. These findings illuminate GluN3A-NMDAR physiology and mechanism, with implications for neuronal signaling and pharmacology.

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Xu, L., de battista, M., Yao, K., Schwenk, J., Nehme, L., pizzamiglio, l., Cerasale, A., Fakler, B., Stroebel, D., Zhu, S., Paoletti, P.. 2026-03-31. Distribution, assembly and mechanism of GluN1/GluN3A excitatory glycine receptors. https://doi.org/10.64898/2026.03.29.715060

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