title: Cellular Protection of SNAP-25 against Botulinum Neurotoxin/A: Inhibition of Thioredoxin Reductase through a Suicide Substrate Mechanism.
abstract: Botulium neurotoxins (BoNTs) are among the most lethal toxins known to man. They are comprised of seven serotypes with BoNT/A being the most deadly; yet, there is no approved therapeutic for their intoxication or one that has even advanced to clinical trials. Botulinum neurotoxicity is ultimately governed through light chain (LC) protease SNARE protein cleavage leading to a loss of neurotransmitter release. Pharmacological attempts to ablate BoNT/A intoxication have sought to either nullify cellular toxin entry or critical biochemical junctions found within its intricate mechanism of action. In these regards, reports have surfaced of nonpeptidic small molecule inhibitors, but few have demonstrated efficacy in neutralizing cellular toxicity, a key prerequisite before rodent lethality studies can be initiated. On the basis of a lead discovered in our BoNT/A cellular assay campaign, we investigated a family of N-hydroxysuccinimide inhibitors grounded upon structure activity relationship (SAR) fundamentals. Molecules stemming from this SAR exercise were theorized to be protease inhibitors. However, this proposition was overturned on the basis of extensive kinetic analysis. Unexpectedly, inhibitor data pointed to thioredoxin reductase (TrxR), an essential component required for BoNT protease translocation. Also unforeseen was the inhibitors' mechanism of action against TrxR, which was found to be brokered through a suicide-mechanism utilizing quinone methide as the inactivating element. This new series of TrxR inhibitors provides an alternative means to negate the etiological agent responsible for BoNT intoxication, the LC protease.
authors.forename: Hajime
authors.initials: H
authors.lastname: Seki
authors.forename: Song
authors.initials: S
authors.lastname: Xue
authors.affiliation: Department of Bacteriology, University of Wisconsin , 1550 Linden Drive, Madison, Wisconsin 53706, United States.
authors.forename: Sabine
authors.initials: S
authors.lastname: Pellett
authors.forename: Peter
authors.initials: P
authors.lastname: Šilhár
authors.affiliation: Department of Bacteriology, University of Wisconsin , 1550 Linden Drive, Madison, Wisconsin 53706, United States.
authors.forename: Eric A
authors.initials: EA
authors.lastname: Johnson
authors.forename: Kim D
authors.initials: KD
authors.lastname: Janda
mesh_terms: D019274:Botulinum Toxins, Type A / Q000037:antagonists & inhibitors / Q000633:toxicity*; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D050825:Synaptosomal-Associated Protein 25 / Q000378:metabolism*; D013880:Thioredoxin-Disulfide Reductase / Q000037:antagonists & inhibitors*
chemical_list: D004791:Enzyme Inhibitors; C494208:SNAP25 protein, human; D050825:Synaptosomal-Associated Protein 25; D013880:Thioredoxin-Disulfide Reductase; D019274:Botulinum Toxins, Type A