Peptides modulating conformational changes in secreted chaperones: From in silico design to preclinical proof of concept
Peptides modulating conformational changes in secreted chaperones: From in silico design to preclinical proof of concept
复制标题
DOI：
10.1073/pnas.0906514106
复制
发表时间：
2009-08-18
影响因子：
11.1
通讯作者：
Borukhov, Itamar
中科院分区：
文献类型：
--
作者：
Kliger, Yossef;Levy, Ofer;Borukhov, Itamar
阻断生物活性蛋白的构象变化具有治疗前景。受合成肽抑制病毒进入的敏感性的启发，合成肽阻断病毒包膜蛋白中螺旋-螺旋相互作用的形成，我们开发了一种预测相互作用螺旋的计算方法。利用这种结合了相关突变分析和傅里叶变换的方法，我们设计了针对gp96和clusterin的肽，这两种分泌的伴侣蛋白已知在非活性构象和活性构象之间转换。在人血单核细胞中，gp96衍生肽对内毒素诱导的TNF α、IL-1 β、IL-6和IL-8的产生抑制作用达80%。当注射到小鼠体内时，该肽将内毒素诱导的TNF α、IL-6和IFN γ的循环水平降低了50%。在体外和肺癌异种移植模型中，簇素衍生的肽抑制了几种肿瘤细胞系的增殖，并显著增强了紫杉醇的细胞抑制活性。实验还验证了所预测的活性肽的作用模式。这两种肽都与它们的亲本蛋白结合，它们的生物活性在与对应螺旋对应的肽存在时被废除。这些数据证明了一种以前未表征的合理设计蛋白质拮抗剂的方法。
Blocking conformational changes in biologically active proteins holds therapeutic promise. Inspired by the susceptibility of viral entry to inhibition by synthetic peptides that block the formation of helix-helix interactions in viral envelope proteins, we developed a computational approach for predicting interacting helices. Using this approach, which combines correlated mutations analysis and Fourier transform, we designed peptides that target gp96 and clusterin, 2 secreted chaperones known to shift between inactive and active conformations. In human blood mononuclear cells, the gp96-derived peptide inhibited the production of TNF alpha, IL-1 beta, IL-6, and IL-8 induced by endotoxin by >80%. When injected into mice, the peptide reduced circulating levels of endotoxin-induced TNF alpha, IL-6, and IFN gamma by >50%. The clusterin-derived peptide arrested proliferation of several neoplastic cell lines, and significantly enhanced the cytostatic activity of taxol in vitro and in a xenograft model of lung cancer. Also, the predicted mode of action of the active peptides was experimentally verified. Both peptides bound to their parent proteins, and their biological activity was abolished in the presence of the peptides corresponding to the counterpart helices. These data demonstrate a previously uncharacterized method for rational design of protein antagonists.