原癌蛋白KRas的表达和纯化及其与小分子抑制剂ZCL1688的分子对接分析

牟榕梓 孙暄 吴更

引用本文: 牟榕梓, 孙暄, 吴更. 原癌蛋白KRas的表达和纯化及其与小分子抑制剂ZCL1688的分子对接分析[J]. 分析化学, 2022, 50(2): 263-270. doi: 10.19756/j.issn.0253-3820.210554 shu
Citation:  MU Rong-Zi,  SUN Xuan,  WU Geng. Expression and Purification of Proto-oncoprotein KRas and Its Molecular Docoking with Small Molecule Inhibitor ZCL1688[J]. Chinese Journal of Analytical Chemistry, 2022, 50(2): 263-270. doi: 10.19756/j.issn.0253-3820.210554 shu

原癌蛋白KRas的表达和纯化及其与小分子抑制剂ZCL1688的分子对接分析

    通讯作者: 吴更,E-mail:geng.wu@sjtu.edu.cn
  • 基金项目:

    国家自然科学基金项目(No.31670106)资助

摘要: 经过特殊设计的小分子抑制剂可共价结合KRas蛋白的催化结构域,从而抑制KRas与下游效应蛋白的结合。本研究采用镍柱亲和层析、酶切标签、凝胶过滤层析等步骤制备了高纯度的KRas催化结构域(氨基酸残基1-169)蛋白;等温滴定量热(ITC)实验证明了KRas蛋白通过K42残基与小分子抑制剂ZCL1688进行结合;通过分子对接得到KRas蛋白与ZCL1688的复合物的建模结构,发现两者的结合对KRas与下游蛋白丝氨酸/苏氨酸激酶RAF的结合有明显竞争作用。本研究建立了一种简便的KRas蛋白催化结构域(氨基酸残基1-169)高效表达与高纯度制备方法,模拟了此蛋白与ZCL1688的结合模式,并分析了该结合模式下的相互作用,为后续筛选此蛋白与小分子抑制剂的共结晶以及复合物结构解析奠定了良好基础。

English


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  • 收稿日期:  2021-06-08
  • 修回日期:  2021-12-16
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