Chemical probes of protein histidine phosphatase activity

项目摘要 蛋白质组氨酸磷酸化在多种关键信号通路中发挥重要作用 但人们对此知之甚少。这种信息匮乏的主要原因是 组氨酸磷酸化是PHIs相对于pSer、pThr和pTyr的相对不稳定 以及完全缺乏可用于监测和调节相关酶的化学工具 组氨酸的磷酸化和去磷酸化。在这个项目中，我们将开发荧光技术 可同时监测组氨酸磷酸酶和组氨酸激酶活性的检测方法 并从一系列重点筛选工作中确定第一批组氨酸磷酸酶抑制剂。 本课题是对PAR-17-046《技术探索性研究》的响应 发展(R21)“，如PAR所示，重点是发展创新的、 研究组氨酸磷酸化和去磷酸化的使能技术。未来 工作的中心将是应用这些工具来更好地理解角色 哺乳动物细胞中的单个组氨酸磷酸酶。

Project Summary Protein histidine phosphorylation plays an important role in several key signaling pathways in mammals but is very poorly understood. The main reasons for this dearth of information about histidine phosphorylation are the relative instability of pHis as compared to pSer, pThr, and pTyr and a complete lack of chemical tools available to monitor and modulate the enzymes involved in histidine phosphorylation and dephosphorylation. In this project, we will develop fluorogenic assays that can be applied to monitoring both histidine phosphatase and histidine kinase activity and identify the first histidine phosphatase inhibitors from a series of focused screening efforts. This project is submitted in response to PAR-17-046 “Exploratory Research for Technology Development (R21)” and, as indicated in the PAR, is focused on the development of innovative, enabling technologies for studying histidine phosphorylation and dephosphorylation. Future work would center around applying these tools to developing a better understanding of the roles individual histidine phosphatases in mammalian cells.