基于均相时间分辨荧光技术(HTRF)的HSP90-HOP相互作用抑制剂活性测试方法的构建及其应用

HTRF-based method for determination of HSP90-HOP inhibition activity and its application

热休克蛋白90(HSP90)是细胞内大量表达的一类蛋白,其主要功能是辅助细胞内其他蛋白(客户蛋白)的成熟。HSP90的众多客户蛋白与肿瘤的发生发展有重要的关系,因此抑制HSP90的功能可以使这些癌症相关蛋白降解,从而诱导肿瘤细胞凋亡,达到治疗肿瘤的目的。HSP90在发挥功能时还需要与辅伴侣蛋白HOP相互作用,因此靶向HSP90-HOP的相互作用开展抑制剂研发成为抑制HSP90伴侣系统的新策略。本文基于均相时间分辨荧光技术(HTRF)构建了稳定的用于测试HSP90-HOP相互作用抑制剂活性的方法,并用该方法研究了HSP90 C-端五肽MEEVD及其突变肽段对HSP90-HOP相互作用的抑制活性,为新型HSP90-HOP相互作用抑制剂的筛选和发现提供了重要基础。

HSP90 is widely expressed in cells with the main function in assisting the maturation of other proteins that are called clients. Many clients play critical roles in the occurrence and development of cancer. Inhibition of HSP90 can lead to degradation of the oncogenic proteins, and result in potent anti-cancer effects. HSP90-HOP interaction is critical for the chaperone function of HSP90, thereby disruption of the HSP90-HOP interaction is a novel strategy in the inhibition of HsPg0. Based on the technology of homogeneous time-resolved fluorescence （HTRF）, we developed a new assay for the identification of new inhibitors of HSP90-HOP interaction. This method was evaluated in the study of the HSP90-HOP inhibition activity of the pentapeptide MEEVD from HSP90 C-terminal and its derivatives. This study can provide a basis for the screening and discovery of novel HSP90-HOP disruptors.