Structural basis for high-voltage activation and subtype-specific inhibition of human Na(v)1.8

The dorsal root ganglia–localized voltage-gated sodium (Na(v)) channel Na(v)1.8 represents a promising target for developing next-generation analgesics. A prominent characteristic of Na(v)1.8 is the requirement of more depolarized membrane potential for activation. Here we present the cryogenic electron microscopy structures of human Na(v)1.8 alone and bound to a selective pore blocker, A-803467, at overall resolutions of 2.7 to 3.2 Å. The first voltage-sensing domain (VSD(I)) displays three different conformations. Structure-guided mutagenesis identified the extracellular interface between VSD(I) and the pore domain (PD) to be a determinant for the high-voltage dependence of activation. A-803467 was clearly resolved in the central cavity of the PD, clenching S6(IV). Our structure-guided functional characterizations show that two nonligand binding residues, Thr397 on S6(I) and Gly1406 on S6(III), allosterically modulate the channel’s sensitivity to A-803467. Comparison of available structures of human Na(v) channels suggests the extracellular loop region to be a potential site for developing subtype-specific pore-blocking biologics.