Wilcox, M. R.; Nigam, A.;Glasgow, N. G.; Narangoda, C.; Phillips, M. B.; Patel, D. S.; Mesbahi-Vasey, S.; Turcu, A. T.; Vázquez, S.; Kurnikova, M. G.; Johnson, J. W. Nat. Commun. 2022, doi: https://www.nature.com/articles/s41467-022-31817-z

N-methyl-d-aspartate receptors (NMDARs) are transmembrane proteins that are activated by the neurotransmitter glutamate and are found at most excitatory vertebrate synapses. NMDAR channel blockers, an antagonist class of broad pharmacological and clinical significance, inhibit by occluding the NMDAR ion channel. A vast literature demonstrates that NMDAR channel blockers, including MK-801, phencyclidine, ketamine, and the Alzheimer’s disease drug memantine, can bind and unbind only when the NMDAR channel is open. Here we use electrophysiological recordings from transfected tsA201 cells and cultured neurons, NMDAR structural modeling, and custom-synthesized compounds to show that NMDAR channel blockers can enter the channel through two routes: in addition to the well-known hydrophilic path from extracellular solution to channel through the open channel gate, a hydrophobic path from plasma membrane to channel through a gated fenestration (“membrane-to-channel inhibition” (MCI)). Our demonstration that ligand-gated channels are subject to MCI, as are voltage-gated channels, highlights the broad significance of this inhibitory mechanism.