Design and biological testing of peptidic dimerization inhibitors of human Hsp90 that target the C-terminal domain
Design and biological testing of peptidic dimerization inhibitors of human Hsp90 that target the C-terminal domain
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DOI：
10.1016/j.bbagen.2016.01.005
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发表时间：
2016-06-01
影响因子：
3
通讯作者：
Jose, Joachim
中科院分区：
文献类型：
--
作者：
Bopp, Bertan;Ciglia, Emanuele;Jose, Joachim
背景：针对癌症治疗有效靶点 Hsp90 C 端结构域二聚化界面的小分子尚未确定。方法：设计了三种肽，旨在抑制 Hsp90 二聚化。计算和生物物理方法检查了三种肽的α螺旋结构。基于Autodisplay技术，开发了一种新型流式细胞仪二聚化检测方法来测试Hsp90二聚化的抑制作用。使用微尺度热泳来确定肽对 Hsp90 C 端结构域的 K-D。结果：MD 模拟和 CD 光谱表明三个肽中的两个具有 α 螺旋结构。通过流式细胞仪分析，测定肽H2的IC50值为2.08μM，肽H3的IC50值为8.96μM。通过微尺度热泳分析 C 末端二聚化结构域的二聚体形成，并确定 K-D 为 129 nM。此外，微尺度热泳研究表明，H2 和 H3 与 C 端结构域具有高亲和力结合，K-D 分别为 1.02 μM 和 1.46 μM。 结论：这些结果揭示了第一个针对 C 端结构域的 Hsp90 二聚化肽抑制剂。此外，研究表明这些肽以低微摩尔亲和力与C端结构域结合。一般意义：这些结果可用于设计和筛选抑制Hsp90 C端结构域二聚化的小分子，从而为癌症治疗开辟新途径。 (C) 2016 年由 Elsevier B.V. 出版
Background: Small molecules targeting the dimerization interface of the C-terminal domain of Hsp90, a validated target for cancer treatment, have yet to be identified.Methods: Three peptides were designed with the aim to inhibit the dimerization of Hsp90. Computational and biophysical methods examined the alpha-helical structure for the three peptides. Based on the Autodisplay technology, a novel flow cytometer dimerization assay was developed to test inhibition of Hsp90 dimerization. Micro scale thermophoresis was used to determine the K-D of the peptides towards the C-terminal domain of Hsp90.Results: MD simulations and CD spectroscopy indicated an alpha-helical structure for two of the three peptides. By flow cytometer analysis, IC50 values of 2.08 mu M for peptide H2 and 8.96 mu M for peptide H3 were determined. Dimer formation of the C-terminal dimerization domain was analyzed by microscale thermophoresis, and a K-D of 129 nM was determined. Furthermore, microscale thermophoresis studies demonstrated a high affinity binding of H2 and H3 to the C-terminal domain, with a K-D of 1.02 mu M and 1.46 mu M, respectively.Conclusions: These results revealed the first peptidic inhibitors of Hsp90 dimerization targeting the C-terminal domain. Furthermore, it has been shown that these peptides bind to the C-terminal domain with a low micromolar affinity.General significance: These results can be used to design and screen for small molecules that inhibit the dimerization of the C-terminal domain of Hsp90, which could open a new route for cancer therapy. (C) 2016 Published by Elsevier B.V.