LAUSR

Significance Pannexin-1 (Panx1) channels contribute to neurological disorders, including stroke and epilepsy, where their function has been linked to N-methyl D-aspartate (NMDA) receptors (NMDARs). We discovered that Ca2+ entry via NMDARs recruits endoplasmic reticulum–resident STIM proteins to activate Panx1 by binding to a hydrophobic region localized to the Panx1 N terminus. Using loss-of-function approaches, combined with molecular replacement and use of a STIM/Panx1 function–blocking antibody, we demonstrate that disrupting the STIM/Panx1 interaction prevents Panx1 activation by NMDARs, but not by hypotonic stimuli. Thus, our findings serve as a basis for the design of modality-specific inhibitors against STIM-dependent Panx1 activation that will aid in understanding the multimodal functions of Panx1 and their contribution to physiology and pathology.