Investigation of Salicylanilides as Botulinum Toxin Antagonists.
Investigation of Salicylanilides as Botulinum Toxin Antagonists.
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DOI：
10.1021/acsinfecdis.2c00230
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发表时间：
2022-08-12
影响因子：
5.3
通讯作者：
Janda, Kim D.
中科院分区：
文献类型：
--
作者：
Patel, Ealin N.;Lin, Lucy;Sneller, Molly M.;Eubanks, Lisa M.;Tepp, William H.;Pellett, Sabine;Janda, Kim D.
A型肉毒神经毒素(BONT/A)是疾控中心公认的最有效的毒素，也是一种一级生物杀伤剂。由BONT/A引起的肉毒杆菌中毒的严重性和持久性是一个重要的治疗问题，早期使用抗毒素抗体治疗是唯一被批准的肉毒杆菌中毒的药物治疗。小分子治疗策略既针对重链(HC)，也针对轻链(LC)催化活性部位和α-/β-Exosite。LC的转运机制也已被研究，但一种有效的、无毒的抑制剂仍处于探索阶段。在这项工作中，我们筛选了水杨胺作为潜在的易位抑制剂的文库。对初筛后的潜在导联进行进一步研究，确定SAL30的细胞IC50值为141 nM。通过DQ-牛血清白蛋白荧光、共聚焦显微镜和V-ATPase抑制实验，探讨了水杨胺盐30‘S的作用机理。综上所述，这些发现表明，通过sal30的原生质体机制进行的内切酶体内质子转移导致内体pH升高，从而阻止LC转移到需要酸性pH的细胞质中。因此，水杨胺对BoNT/A活性的抑制可能是通过破坏依赖于pH的内体LC易位来实现的。我们进一步探讨了SAL30对BONT/A LC转位抑制剂巴菲罗霉素A1的抑制作用，这表明两个离子载体之间没有协同作用。
Botulinum neurotoxin serotype A (BoNT/A) is recognized by the CDC as the most potent toxin and as a Tier 1 biowarfare agent. The severity and longevity of botulism stemming from BoNT/A is of significant therapeutic concern and early administration of antitoxin-antibody therapy is the only approved pharmaceutical treatment for botulism. Small molecule therapeutic strategies have targeted both the heavy chain (HC), and the light chain (LC) catalytic active site and α-/β-exosites. The LC translocation mechanism has also been studied, but an effective, non-toxic inhibitor remains under-explored. In this work, we screened a library of salicylanilides as potential translocation inhibitors. Potential leads following a primary screen were further scrutinized identifying sal30 which has a cellular IC50 value of 141 nM. Inquiry of salicylanilide sal30’s mechanism of action was explored through DQ-BSA fluorescence, confocal microscopy, and V-ATPase inhibition assays. The summation of these findings imply that endo-lysosomal proton translocation through the protonophore mechanism of sal30 causes endosome pH to increase, that in turn prevents LC translocation into cytosol which requires acidic pH. Thus, inhibition of BoNT/A activity by salicylanilides likely occurs through disruption of pH-dependent endosomal LC translocation. We further probed BoNT inhibition by sal30 using additivity analysis studies with bafilomycin A1, a known BoNT/A LC translocation inhibitor, which indicated the absence of synergy between the two ionophores.
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影响因子：
4.8
作者：
Chen W;Mook RA Jr;Premont RT;Wang J
通讯作者：
Wang J
影响因子：
4.2
作者：
Ruthel G;Burnett JC;Nuss JE;Wanner LM;Tressler LE;Torres-Melendez E;Sandwick SJ;Retterer CJ;Bavari S
通讯作者：
Bavari S
影响因子：
7.3
作者：
Blake, Steven;Thanissery, Rajani;Janda, Kim D.
通讯作者：
Janda, Kim D.
双质子载体-几丁质酶抑制剂显着影响盘旋圆线虫的蜕皮。
影响因子：
7.3
作者：
Gooyit M;Tricoche N;Lustigman S;Janda KD
通讯作者：
Janda KD
影响因子：
4.6
作者：
Czaplinska, Barbara;Malarz, Katarzyna;Musiol, Robert
通讯作者：
Musiol, Robert