Development of artificial receptors that can recognize a specific protein surface in aqueous solution

开发能够识别水溶液中特定蛋白质表面的人工受体

基本信息

批准号：
15350104
负责人：
HAMACHI Itaru
金额：
$ 9.6万
依托单位：
Kyoto University (2005)Kyushu University (2003-2004)
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

During the project term, molecular recognition chemistry toward phosphorylated proteins and peptides has greatly advanced both in fundamental science and application. My group newly developed artificial receptor molecules based on Zn-metal complexes which can recognize phosphorylated peptides and proteins and sense them by a fluorescence signal change. These can be carried out in aqueous solution and SDS-PAGE gel plates. Detailed study using isothermal titration calorimetry clearly demonstrates that the Zn sites of the receptors tightly bind to the anionic phospate sites of the peptide and thus shows the strong affinity. Further more, using this strong binding, we also tested the inhibition capability of the receptors for the phospho-protein protein interactions. In the case of the cross-linking binding mode of the dinuclear Zn complex type of receptors, it is clear that the effective disruption of the phospho-protein protein interactions between WW domain and the phosphorylated C-terminal domain of Polymerase. These results clearly indicate that artificial receptors can act not only as a simple fluorescent chemosensor for phosphorylated peptides and proteins, but also as a poteintial disruptor for the phospho-protein protein interactions. It is expected that the present accomplishment strongly stimulate the relevant research and may lead to a more sophisticated molecule in the near future.

在项目期间，针对磷酸化蛋白质和肽的分子识别化学在基础科学和应用方面都取得了巨大进步。我的小组新开发了基于锌金属复合物的人工受体分子，它可以识别磷酸化的肽和蛋白质，并通过荧光信号变化来感知它们。这些可以在水溶液和 SDS-PAGE 凝胶板上进行。使用等温滴定量热法的详细研究清楚地表明，受体的 Zn 位点与肽的阴离子磷酸盐位点紧密结合，因此显示出很强的亲和力。此外，利用这种强结合，我们还测试了受体对磷蛋白相互作用的抑制能力。在双核Zn复合物类型受体的交联结合模式的情况下，很明显，有效破坏了WW结构域和聚合酶的磷酸化C端结构域之间的磷酸化蛋白质相互作用。这些结果清楚地表明，人工受体不仅可以充当磷酸化肽和蛋白质的简单荧光化学传感器，而且还可以充当磷酸化蛋白质相互作用的潜在破坏者。预计目前的成就将强烈刺激相关研究，并可能在不久的将来产生更复杂的分子。