Targeting Bacillus anthracis toxicity with a genetically selected inhibitor of the PA/CMG2 protein-protein interaction.
Targeting Bacillus anthracis toxicity with a genetically selected inhibitor of the PA/CMG2 protein-protein interaction.
复制标题
DOI：
10.1038/s41598-017-03253-3
复制
发表时间：
2017-06-08
影响因子：
4.6
通讯作者：
Tavassoli A
中科院分区：
文献类型：
--
作者：
Male AL;Forafonov F;Cuda F;Zhang G;Zheng S;Oyston PCF;Chen PR;Williamson ED;Tavassoli A
人CMG 2受体与炭疽杆菌保护性抗原（PA）之间的蛋白质-蛋白质相互作用对于炭疽致死毒素和水肿毒素转运到人细胞中是必不可少的。我们使用遗传编码的高通量筛选平台来筛选320万个环六肽的SICLOPPS文库，以寻找这种蛋白质-蛋白质相互作用的抑制剂。不寻常的是，前3个命中都在文库的随机化区域中含有终止密码子，导致线性肽而不是环状肽。这些肽在体外破坏靶向相互作用;两个通过结合CMG 2起作用，而一个结合PA。通过掺入非天然苯丙氨酸类似物，最有效的CMG 2结合抑制剂的功效得到改善。基于细胞的测定表明，优化的抑制剂保护巨噬细胞免受致死因子的毒性。
The protein-protein interaction between the human CMG2 receptor and the Bacillus anthracis protective antigen (PA) is essential for the transport of anthrax lethal and edema toxins into human cells. We used a genetically encoded high throughput screening platform to screen a SICLOPPS library of 3.2 million cyclic hexapeptides for inhibitors of this protein-protein interaction. Unusually, the top 3 hits all contained stop codons in the randomized region of the library, resulting in linear rather than cyclic peptides. These peptides disrupted the targeted interaction in vitro; two act by binding to CMG2 while one binds PA. The efficacy of the most potent CMG2-binding inhibitor was improved through the incorporation of non-natural phenylalanine analogues. Cell based assays demonstrated that the optimized inhibitor protects macrophages from the toxicity of lethal factor.