Arbuckle, K. Evolutionary context of venom in animals. Evol. Venom. Anim. Toxins 24, 3–31 (2017).
Huang, J., Pan, X. & Yan, N. Structural biology and molecular pharmacology of voltage-gated ion channels. Nat. Rev. Mol. Cell Biol. 25, 904–925 (2024).
de Lera Ruiz, M. & Kraus, R. L. Voltage-gated sodium channels: structure, function, pharmacology, and clinical indications. J. Med. Chem. 58, 7093–7118 (2015).
Armstrong, C. M. Na channel inactivation from open and closed states. Proc. Natl Acad. Sci. USA 103, 17991–17996 (2006).
Chanda, B. & Bezanilla, F. Tracking voltage-dependent conformational changes in skeletal muscle sodium channel during activation. J. Gen. Physiol. 120, 629–645 (2002).
Lazarovici, P. Snake-and spider-venom-derived toxins as lead compounds for drug development. Methods Mol. Biol. https://doi.org/10.1007/978-1-4939-9845-6_1 (2020).
Kalia, J. et al. From foe to friend: using animal toxins to investigate ion channel function. J. Mol. Biol. 427, 158–175 (2015).
Luz Morales-Lazaro, S., Hernández-García, E., Serrano-Flores, B. & Rosenbaum, T. Organic toxins as tools to understand ion channel mechanisms and structure. Curr. Top. Med. Chem. 15, 581–603 (2015).
Escoubas, P. & Bosmans, F. Spider peptide toxins as leads for drug development. Expert Opin. Drug Discov. 2, 823–835 (2007).
Gilchrist, J. & Bosmans, F. Using voltage-sensor toxins and their molecular targets to investigate NaV1.8 gating. J. Physiol. 596, 1863–1872 (2018).
Fan, X. et al. Phrixotoxin-3 binds to three distinct antagonistic sites on human Nav1.6. Cell Res. 35, 610–613 (2025).
Schaller, K. L., Krzemien, D. M., Yarowsky, P. J., Krueger, B. K. & Caldwell, J. H. A novel, abundant sodium channel expressed in neurons and glia. J. Neurosci. 15, 3231–3242 (1995).
Freeman, S. A., Desmazières, A., Fricker, D., Lubetzki, C. & Sol-Foulon, N. Mechanisms of sodium channel clustering and its influence on axonal impulse conduction. Cell. Mol. Life Sci. 73, 723–735 (2016).
Caldwell, J. H., Schaller, K. L., Lasher, R. S., Peles, E. & Levinson, S. R. Sodium channel Nav1.6 is localized at nodes of Ranvier, dendrites, and synapses. Proc. Natl Acad. Sci. USA 97, 5616–5620 (2000).
Hu, W. et al. Distinct contributions of Nav1.6 and Nav1.2 in action potential initiation and backpropagation. Nat. Neurosci. 12, 996–1002 (2009).
O’Brien, J. E. & Meisler, M. H. Sodium channel SCN8A (Nav1.6): properties and de novo mutations in epileptic encephalopathy and intellectual disability. Front. Genet. 4, 213 (2013).
Liu, Y. et al. Neuronal mechanisms of mutations in SCN8A causing epilepsy or intellectual disability. Brain 142, 376–390 (2019).
Blanchard, M. G. et al. De novo gain-of-function and loss-of-function mutations of SCN8A in patients with intellectual disabilities and epilepsy. J. Med. Genet. 52, 330–337 (2015).
Sole, L., Wagnon, J. L. & Tamkun, M. M. Functional analysis of three Nav1.6 mutations causing early infantile epileptic encephalopathy. Biochim. Biophys. Acta 1866, 165959 (2020).
Tanaka, B. S. et al. A gain-of-function mutation in Nav1.6 in a case of trigeminal neuralgia. Mol. Med. 22, 338–348 (2016).
Wagnon, J. L. et al. Loss-of-function variants of SCN8A in intellectual disability without seizures. Neurol. Genet. 3, e170 (2017).
Wagnon, J. L. et al. Partial loss-of-function of sodium channel SCN8A in familial isolated myoclonus. Hum. Mutat. 39, 965–969 (2018).
Samanta, D. et al. Current and emerging precision therapies for developmental and epileptic encephalopathies. Pediatr. Neurol. 168, 67–81 (2025).
Tchaicha, S. et al. Areas of research priorities in epilepsy: a position paper of the European Reference Network for Rare and Complex Epilepsies, EpiCARE. Epilepsia Open 11, 291–321 (2026).
Pan, X. et al. Molecular basis for pore blockade of human Na+ channel Nav1.2 by the μ-conotoxin KIIIA. Science 363, 1309–1313 (2019).
Shen, H., Liu, D., Wu, K., Lei, J. & Yan, N. Structures of human Nav1.7 channel in complex with auxiliary subunits and animal toxins. Science 363, 1303–1308 (2019).
Huang, G. et al. High-resolution structures of human Nav1.7 reveal gating modulation through α-π helical transition of S6IV. Cell Rep. 39, 110735 (2022).
Li, Y. et al. Structure of human NaV1. 6 channel reveals Na+ selectivity and pore blockade by 4,9-anhydro-tetrodotoxin. Nat. Commun. 14, 1030 (2023).
Wu, T., Yang, X., Jin, X., Yan, N. & Li, Z. Critical role of extracellular loops in differential modulations of TTX-sensitive and TTX-resistant Nav channels. Proc. Natl Acad. Sci. USA 122, e2510355122 (2025).
Shen, H. et al. Structural basis for the modulation of voltage-gated sodium channels by animal toxins. Science 362, eaau2596 (2018).
Clairfeuille, T. et al. Structural basis of α-scorpion toxin action on Nav channels. Science 363, eaav8573 (2019).
Jiang, D. et al. Structural basis for voltage-sensor trapping of the cardiac sodium channel by a deathstalker scorpion toxin. Nat. Commun. 12, 128 (2021).
Fiedler, B. et al. Specificity, affinity and efficacy of iota-conotoxin RXIA, an agonist of voltage-gated sodium channels Nav1.2, 1.6 and 1.7. Biochem. Pharmacol. 75, 2334–2344 (2008).
Jimenez, E. C. et al. Novel excitatory Conus peptides define a new conotoxin superfamily. J. Neurochem. 85, 610–621 (2003).
Schiavon, E. et al. Resurgent current and voltage sensor trapping enhanced activation by a β-scorpion toxin solely in Nav1.6 channel significance in mice Purkinje neurons. J. Biol. Chem. 281, 20326–20337 (2006).
Aili, S. R. et al. An integrated proteomic and transcriptomic analysis reveals the venom complexity of the bullet ant Paraponera clavata. Toxins 12, 324 (2020).
Buczek, O. et al. Structure and sodium channel activity of an excitatory I1-superfamily conotoxin. Biochemistry 46, 9929–9940 (2007).
Buczek, O., Yoshikami, D., Bulaj, G., Jimenez, E. C. & Olivera, B. M. Post-translational amino acid isomerization: a functionally important d-amino acid in an excitatory peptide. J. Biol. Chem. 280, 4247–4253 (2005).
Piek, T. et al. Poneratoxin, a novel peptide neurotoxin from the venom of the ant, Paraponera clavata. Compar. Biochem. Physiol. C 99, 487–495 (1991).
Robinson, S. D. et al. Ant venoms contain vertebrate-selective pain-causing sodium channel toxins. Nat. Commun. 14, 2977 (2023).
Fan, X., Huang, J., Jin, X. & Yan, N. Cryo-EM structure of human voltage-gated sodium channel Nav1.6. Proc. Natl Acad. Sci. USA 120, e2220578120 (2023).
Wu, Q. et al. Structural mapping of Nav1.7 antagonists. Nat. Commun. 14, 3224 (2023).
Shen, H. et al. Structural basis for the modulation of voltage-gated sodium channels by animal toxins. Science https://doi.org/10.1126/science.aau2596 (2018).
Huang, G. et al. Unwinding and spiral sliding of S4 and domain rotation of VSD during the electromechanical coupling in Nav1.7. Proc. Natl Acad. Sci. USA 119, e2209164119 (2022).
Szolajska, E. et al. Poneratoxin, a neurotoxin from ant venom: structure and expression in insect cells and construction of a bio-insecticide. Eur. J. Biochem. 271, 2127–2136 (2004).
Fan, X. et al. Open-state structure of veratridine-activated human Nav1.7 reveals the molecular choreography of fast inactivation. Vita https://doi.org/10.15302/vita.2026.01.0003 (2026).
Jin, X., Huang, J., Wang, H., Wang, K. & Yan, N. A versatile residue numbering scheme for Nav and Cav channels. Cell Chem. Biol. 31, 1394–1404 (2024).
Israel, M. R. et al. The E15R point mutation in scorpion toxin Cn2 uncouples its depressant and excitatory activities on human Nav1.6. J. Med. Chem. 61, 1730–1736 (2018).
Jiang, D. et al. Open-state structure and pore gating mechanism of the cardiac sodium channel. Cell 184, 5151–5162 (2021).
Wisedchaisri, G. & Gamal El-Din, T. M. Druggability of voltage-gated sodium channels-exploring old and new drug receptor sites. Front. Pharmacol. 13, 858348 (2022).
Fattorusso, A. et al. The pharmacoresistant epilepsy: an overview on existant and new emerging therapies. Front. Neurol. 12, 674483 (2021).
Stevens, M., Peigneur, S. & Tytgat, J. Neurotoxins and their binding areas on voltage-gated sodium channels. Front. Pharmacol. 2, 71 (2011).
Bagal, S. K., Marron, B. E., Owen, R. M., Storer, R. I. & Swain, N. A. Voltage gated sodium channels as drug discovery targets. Channels 9, 360–366 (2015).
Blumenfeld, H. et al. Role of hippocampal sodium channel Nav1.6 in kindling epileptogenesis. Epilepsia 50, 44–55 (2009).
Phulera, S. et al. Scorpion α-toxin LqhαIT specifically interacts with a glycan at the pore domain of voltage-gated sodium channels. Structure https://doi.org/10.1016/j.str.2024.07.021 (2024).
Cui, C. et al. Mechanisms of KCNQ1 gating modulation by KCNE1/3 for cell-specific function. Cell Res. 35, 876–886 (2025).
Goldschen-Ohm, M. P., Capes, D. L., Oelstrom, K. M. & Chanda, B. Multiple pore conformations driven by asynchronous movements of voltage sensors in a eukaryotic sodium channel. Nat. Commun. 4, 1350 (2013).
Gao, S. et al. Structural basis for human Cav1.2 inhibition by multiple drugs and the neurotoxin calciseptine. Cell 186, 5363–5374 (2023).
Gao, S., Yao, X. & Yan, N. Structure of human Cav2.2 channel blocked by the painkiller ziconotide. Nature 596, 143–147 (2021).
Pettersen, E. F. et al. UCSF ChimeraX: structure visualization for researchers, educators, and developers. Protein Sci. 30, 70–82 (2021).
Smart, O. S., Neduvelil, J. G., Wang, X., Wallace, B. A. & Sansom, M. S. HOLE: a program for the analysis of the pore dimensions of ion channel structural models. J. Mol. Graph. 14, 354–360 (1996).
Punjani, A., Rubinstein, J. L., Fleet, D. J. & Brubaker, M. A. cryoSPARC: algorithms for rapid unsupervised cryo-EM structure determination. Nat. Methods 14, 290–296 (2017).
Lei, J. & Frank, J. Automated acquisition of cryo-electron micrographs for single particle reconstruction on an FEI Tecnai electron microscope. J. Struct. Biol. 150, 69–80 (2005).
Bepler, T. et al. Positive-unlabeled convolutional neural networks for particle picking in cryo-electron micrographs. Nat. Methods 16, 1153–1160 (2019).
Emsley, P., Lohkamp, B., Scott, W. G. & Cowtan, K. Features and development of Coot. Acta Crystallogr. D 66, 486–501 (2010).
Pintar, A., Possani, L. D. & Delepierre, M. Solution structure of toxin 2 from Centruroides noxius Hoffmann, a β-scorpion neurotoxin acting on sodium channels. J. Mol. Biol. 287, 359–367 (1999).
Afonine, P. V. et al. Towards automated crystallographic structure refinement with phenix.refine. Acta Crystallogr. D 68, 352–367 (2012).
Croll, T. I. ISOLDE: a physically realistic environment for model building into low-resolution electron-density maps. Acta Crystallogr. D 74, 519–530 (2018).
Abramson, J. et al. Accurate structure prediction of biomolecular interactions with AlphaFold 3. Nature 630, 493–500 (2024).
Li, Z. et al. Structure of human Nav1.5 reveals the fast inactivation-related segments as a mutational hotspot for the long QT syndrome. Proc. Natl Acad. Sci. USA 118, e2100069118 (2021).
Huang, X. et al. Structural basis for high-voltage activation and subtype-specific inhibition of human Nav1.8. Proc. Natl Acad. Sci. USA 119, e2208211119 (2022).
Sastry, G. M., Adzhigirey, M., Day, T., Annabhimoju, R. & Sherman, W. Protein and ligand preparation: parameters, protocols, and influence on virtual screening enrichments. J. Comput. Aided Mol. Des. 27, 221–234 (2013).
Lu, C. et al. OPLS4: improving force field accuracy on challenging regimes of chemical space. J. Chem. Theory Comput. 17, 4291–4300 (2021).
Lomize, A. L., Todd, S. C. & Pogozheva, I. D. Spatial arrangement of proteins in planar and curved membranes by PPM 3.0. Protein Sci. 31, 209–220 (2022).
Mark, P. & Nilsson, L. Structure and dynamics of the TIP3P, SPC, and SPC/E water models at 298 K. J. Phys. Chem. A 105, 9954–9960 (2001).
McGibbon, R. T. et al. MDTraj: a modern open library for the analysis of molecular dynamics trajectories. Biophys. J. 109, 1528–1532 (2015).
Pérez-Hernández, G. & Hildebrand, P. W. mdciao: accessible analysis and visualization of molecular dynamics simulation data. PLoS Comput. Biol. 21, e1012837 (2025).
