Conotoxins are disulfide-rich peptides with exquisite pharmacological potency and selectivity across diverse protein targets. ⍺-conotoxins (⍺-CTX) selectively inhibit both neuronal and muscle-type nicotinic acetylcholine receptors (nAChR) with remarkable subtype specificity, which makes them promising molecules for the development of novel therapeutics. The underlying molecular determinants of this specificity have remained elusive, and studies have been hindered by the absence of structural insights into the receptor-toxin interface. Here, we present the first high-resolution structures of ⍺-CTX bound to full-length nAChR and begin to parse the precise pharmacophore drivers of potency and selectivity. Our integrated approach combines structural studies with pharmacological profiling and computational modelling, which allowed us to identify conserved moieties that govern nAChR recognition and inhibition. This work establishes a framework for the rational development of conotoxin-based peptides and conotoxin based blocking agents.