title: Mutant cycle analysis with modified saxitoxins reveals specific interactions critical to attaining high-affinity inhibition of hNaV1.7.
abstract: Improper function of voltage-gated sodium channels (NaVs), obligatory membrane proteins for bioelectrical signaling, has been linked to a number of human pathologies. Small-molecule agents that target NaVs hold considerable promise for treatment of chronic disease. Absent a comprehensive understanding of channel structure, the challenge of designing selective agents to modulate the activity of NaV subtypes is formidable. We have endeavored to gain insight into the 3D architecture of the outer vestibule of NaV through a systematic structure-activity relationship (SAR) study involving the bis-guanidinium toxin saxitoxin (STX), modified saxitoxins, and protein mutagenesis. Mutant cycle analysis has led to the identification of an acetylated variant of STX with unprecedented, low-nanomolar affinity for human NaV1.7 (hNaV1.7), a channel subtype that has been implicated in pain perception. A revised toxin-receptor binding model is presented, which is consistent with the large body of SAR data that we have obtained. This new model is expected to facilitate subsequent efforts to design isoform-selective NaV inhibitors.
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305.
authors.forename: Rhiannon
authors.initials: R
authors.lastname: Thomas-Tran
authors.affiliation: Department of Chemistry, Stanford University, Stanford, CA 94305 jdubois@stanford.edu.
authors.forename: J
authors.initials: J
authors.lastname: Du Bois
mesh_terms: D000818:Animals; D001665:Binding Sites; D016466:CHO Cells; D003412:Cricetulus; D015195:Drug Design; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D062105:Molecular Docking Simulation; D009124:Muscle Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D009154:Mutation; D062551:NAV1.2 Voltage-Gated Sodium Channel / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D062554:NAV1.5 Voltage-Gated Sodium Channel / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D062556:NAV1.7 Voltage-Gated Sodium Channel / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011485:Protein Binding; D011487:Protein Conformation; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012530:Saxitoxin / Q000737:chemistry / Q000494:pharmacology*; D026941:Sodium Channel Blockers / Q000737:chemistry / Q000494:pharmacology*; D015222:Sodium Channels / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D013779:Tetrodotoxin / Q000737:chemistry / Q000494:pharmacology
chemical_list: D009124:Muscle Proteins; D062551:NAV1.2 Voltage-Gated Sodium Channel; D062554:NAV1.5 Voltage-Gated Sodium Channel; D062556:NAV1.7 Voltage-Gated Sodium Channel; D011994:Recombinant Proteins; C568320:SCN5A protein, human; C486866:SCN9A protein, human; C568253:Scn2A protein, rat; C466180:Scn4a protein, rat; D026941:Sodium Channel Blockers; D015222:Sodium Channels; D012530:Saxitoxin; D013779:Tetrodotoxin
keywords: guanidinium toxin; mutant cycle analysis; sodium channel