A genetically encoded secreted toxin potentiates synaptic NMDA receptors in hippocampal neurons and confers neuroprotection

NMDA receptors (NMDARs) play essential roles in neuronal development, survival, and synaptic plasticity, to name a few. However, dysregulation in receptors' activity can lead to neuronal and synaptic damage, contributing to the development of various brain pathologies. Current pharmacological treatments targeting NMDARs remain limited, for instance due to insufficient receptor selectivity and poor spatial targeting. Genetic approaches hold promise to overcome some of these issues; however, require genetically encodable NMDAR-modulating peptides, which are scarce. Here, we explored NMDAR-selective peptide toxins from marine cone snails, which resulted in the necessary engineering of a posttranslational modification-free variant of Conantokin-P (<i>naked</i> Con-P). The <i>naked</i> form is essential for expression in mammalian cells. We systematically explored the <i>naked</i> variant and discovered that <i>naked</i> Con-P maintains its ability to inhibit GluN2B-containing receptors, but uniquely acquired the ability to potentiate GluN2A-containing synaptic receptors. We then engineered a secreted <i>naked</i> Con-P that readily enhances NMDAR-mediated synaptic events in primary hippocampal neurons, and mitigates neuronal damage induced by staurosporine. We therefore provide a genetically encodable, subtype selective, and secreted bimodulator of NMDARs. This new variant and approach should pave the way for the development of additional genetic tools, specifically tailored to target NMDARs within distinct cellular populations in the brain.