Scorpion α-toxin LqhαIT specifically interacts with a glycan at the pore domain of voltage-gated sodium channels

Abstract

AbstractVoltage-gated sodium (Nav) channels sense membrane potential and drive cellular electrical activity. Numerous protein toxins have been identified that modulate Nav gating, and structures of Nav channels in complex with these toxins helped elucidate the molecular mechanisms of voltage-dependent channel gating. The deathstalker scorpion α-toxin LqhαIT exerts a strong action potential prolonging effect on Nav channels. Biochemical studies show that LqhαIT features a functionally essential epitope at its C-terminus that is not shared with related scorpion α-toxins. To elucidate the mechanism of action of LqhαIT, we determined a 3.9 Å cryo-electron microscopy (cryo-EM) structure of LqhαIT in complex with the Nav channel fromPeriplaneta americana(NavPas). We found that LqhαIT binds to voltage sensor domain 4 and traps it in a “S4 down” conformation to stabilize the open state. To promote binding, the functionally essential C-terminal epitope of LqhαIT forms an extensive interface with the glycan scaffold linked to Asn330 of NavPas that augments a small protein-protein interface between NavPas and LqhαIT. A combination of molecular dynamics simulations, structural comparisons, and prior mutagenesis experiments demonstrate the functional importance of this toxin-glycan interaction. These findings help establish a structural basis for the specificity achieved by scorpion α-toxins and provide crucial insights for the development and optimization of new Nav channel modulators.