Mechanism of differential control of NMDA receptor activity by NR2 subunits

NMDA receptors (NMDARs) are a major class of excitatory neurotransmitter receptors in the central nervous system. They form glutamate-gated ion channels highly permeable to calcium that mediate activity-dependent synaptic plasticity1. NMDAR dysfunction is implicated in multiple brain disorders, including stroke, chronic pain and schizophrenia2. NMDARs exist as multiple subtypes with distinct pharmacological and biophysical properties largely determined by the type of NR2 subunit (NR2A-NR2D) incorporated in the heteromeric NR1/NR2 complex1,3,4. A fundamental difference between NMDAR subtypes is their channel maximal open probability (Po), which spans a 50-fold range from ~0.5 for NR2A-containing receptors to ~0.01 for NR2C- and NR2D-containing receptors; NR2B-containing receptors having an intermediate value (~0.1)5–9. These differences in Po confer unique charge transfer capacities and signaling properties on each receptor subtype4,6,10,11. The molecular basis for this profound difference in activity between NMDAR subtypes is unknown. Here we demonstrate that the subunit-specific gating of NMDARs is controlled by the region formed by the NR2 N-terminal domain (NTD), an extracellular clamshell-like domain previously shown to bind allosteric inhibitors12–15, and the short linker connecting the NTD to the agonist-binding domain (ABD). Subtype specificity of NMDAR Po largely reflects differences in the spontaneous (ligand-independent) equilibrium between open-cleft and closed-cleft conformations of the NR2-NTD. This NTD-driven gating control also impacts pharmacological properties, by setting the sensitivity to the endogenous inhibitors zinc and protons. Our results provide a proof-of-concept for a drug-based bidirectional control of NMDAR activity using molecules acting either as NR2-NTD “closers” or “openers” promoting receptor inhibition or potentiation, respectively.