In Silico Identification and Experimental Validation of Peptide-Based Inhibitors Targeting Clostridium difficile Toxin A
In Silico Identification and Experimental Validation of Peptide-Based Inhibitors Targeting Clostridium difficile Toxin A
复制标题
靶向艰难梭菌毒素 A 的肽抑制剂的计算机模拟鉴定和实验验证
DOI：
10.1021/acschembio.1c00743
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发表时间：
2022
影响因子：
4
通讯作者：
Hall, Carol K.
中科院分区：
文献类型：
--
作者：
Xiao, Xingqing;Sarma, Sudeep;Menegatti, Stefano;Crook, Nathan;Magness, Scott T.;Hall, Carol K.
艰难梭菌感染由两种主要外毒素介导：毒素A（TcdA）和B（TcdB）。用靶向肽类药物抑制这些毒素的生物催化活性可以降低C.艰难感染。在这项工作中，我们使用了一种计算策略，集成了肽结合设计（PepBD）算法和显式溶剂原子分子动力学模拟，以确定有前途的毒素A靶向肽，可以识别和结合到TcdA葡萄糖基转移酶结构域（GTD）的催化位点。模拟结果显示，三种电子发现的肽中有两种，即中和肽A（NPA）和中和肽B（NPB），与TcdA GTD结合时的结合自由能比噬菌体展示肽（RP）低。这些肽可能作为抗C的潜在抑制剂。艰难感染。在体外在人空肠细胞中测试肽RP、NPA和NPB中和TcdA的细胞病变效应的功效。发现噬菌体展示肽RP和硅肽NPA都表现出强的毒素中和特性，从而防止TcdA毒性。然而，硅肽NPB对TcdA表现出相对低的功效。
Clostridium difficileinfection is mediated by two major exotoxins: toxins A (TcdA) and B (TcdB). Inhibiting the biocatalytic activities of these toxins with targeted peptide-based drugs can reduce the risk ofC. difficileinfection. In this work, we used a computational strategy that integrates a peptide binding design (PepBD) algorithm and explicit-solvent atomistic molecular dynamics simulation to determine promising toxin A-targeting peptides that can recognize and bind to the catalytic site of the TcdA glucosyltransferase domain (GTD). Our simulation results revealed that two out of threein silicodiscovered peptides,viz.the neutralizing peptides A (NPA) and B (NPB), exhibit lower binding free energies when bound to the TcdA GTD than the phage-display discovered peptide,viz.the reference peptide (RP). These peptides may serve as potential inhibitors againstC. difficileinfection. The efficacy of the peptides RP, NPA, and NPB to neutralize the cytopathic effects of TcdA was testedin vitroin human jejunum cells. Both phage-display peptide RP andin silicopeptide NPA were found to exhibit strong toxin-neutralizing properties, thereby preventing the TcdA toxicity. However, thein silicopeptide NPB demonstrates a relatively low efficacy against TcdA.