Bidentate Molecular Probes for Hect. E3 Ubiquitin Ligase Biological Function

蛋白质泛素化是一种翻译后修饰，调节多种细胞过程，包括蛋白质周转、运输、亚细胞定位、转录功能和DNA修复机制。泛素偶联是指由泛素激活酶(E1)、泛素偶联酶(E2)和泛素连接酶(E3)依次催化的一系列事件。该项目将致力于开发新的二价分子探针，以阐明Hect E3泛素连接酶这一定义明确的小亚群的生物学作用和底物特异性功能。特别感兴趣的是类似NEDD4的E3泛素蛋白连接酶WWP1和WWP2。后者与转化生长因子信号通路中的SMAD蛋白结合，目标是降解它们。先进的细胞/分子生物学和合成化学的领先申请者的综合优势，加上新思想和新学科的交叉培养，将促进新的方法来阐明基本蛋白质泛素化机制的动力学。使用简单的化学工具来探索复杂的生化途径/网络的单个成分的功能和重要性显然符合BBSRC的职权范围。学生将采用一系列技术(化学合成、机械酶学、全细胞研究)来探索不同配体在细胞和分子水平上的功能。这是一个与BBSRC在化学-生物学界面的研究优先事项相关的主题领域。该项目的成果还可能导致下一代细胞生长的选择性抑制剂，这将对进一步研究和开发生产改进的癌症化疗药物产生极大的兴趣。支持可能在知识转移、商业化和医疗福利方面产生有益影响的研究)是BBSRC越来越重要的优先事项。

Protein ubiquitination represents a post-translational modification that regulates a variety of cellular processes including protein turnover, trafficking, sub-cellular localisation, transcriptional function and DNA repair mechanisms. Ubiquitin conjugation involves a cascade of events catalysed sequentially by E1 (ubiquitin-activating), E2 (ubiquitin conjugating), and E3 (ubiquitin ligase) enzymes. This project will focus on the development of novel bivalent molecular probes to elucidate the biological roles and substrate-specific function of a small well-defined sub-group known as the HECT E3 ubiquitin ligases. Of particular interest will be the NEDD4-like E3 ubiquitin-protein ligases WWP1 and WWP2. The latter binds to SMAD proteins in the TGF- signalling pathway, targeting them for degradation. The combined strengths of the lead applicants in advanced Cell/Molecular Biology and Synthetic Chemistry, together with cross-fertilization of new ideas and disciplines, will facilitate novel approaches to elucidate the dynamics of fundamental protein ubiquitination mechanisms. The use of simple chemical tools to probe the function and importance of individual components of complex biochemical pathways/networks clearly fits within the BBSRC remit. The student will embrace a range of techniques (chemical synthesis, mechanistic enzymology, whole cell studies) to probe the functionality of different ligands at both the cellular and molecular level. This is a subject area relevant to BBSRC research priorities at the Chemistry-Biology interface. The outputs of this project could also lead to the next generation of selective inhibitors of cell growth, which would be of great interest for further study and development to produce improved cancer chemotherapeutics. Supporting research that is likely to have a beneficial impact in terms of knowledge transfer, commercialisation and healthcare benefits) is an increasingly important priority within BBSRC.