Mechanistic understanding and inhibitor design strategies for non-mevalonate pathway TPP-dependent enzymes

非甲羟戊酸途径 TPP 依赖性酶的机理理解和抑制剂设计策略

• 批准号: 2878051
• 金额: --
• 依托单位: University of Leeds
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2878051
• 关键词: Mechanistic; understanding; inhibitor; design; strategies

In this 4-year BBSRC iCASE PhD studentship between the University of Leeds and Syngenta PLC, the student will investigate the structure, function and mechanism of plant enzymes important to the agriscience industry. The project, which is co-supervised in the Astbury Centre for Structural Molecular Biology, University of Leeds by Profs Alex Breeze and Frank Sobott, builds on previous collaboration between Syngenta and the University of Leeds to further elucidate the mechanistic biology and modes of inhibition ofnon-mevalonate pathway TPP-dependent enzymes including 1-deoxy-D-xylulose-5-phosphate synthase (DXS). The outcome will be an enhanced understanding of potential inhibitor design strategies and mechanisms, that in turn may lead to the development of improved herbicidal agents targeting this pathway.Despite belonging to a generally well-understood class of enzymes, mechanistic understanding of DXS and its inhibition is still relatively limited, at least in part because structure determination, particularly of substrate-, catalytic intermediate-, product- or inhibitor-bound forms by X-ray crystallography has proved to be quite challenging. The student will use cutting-edge ultra-high field methyl NMR and hydrogen-deuteriumexchange mass spectrometry (HDX-MS) to build on our earlier work. Specifically, we willtransfer our methyl NMR approach based on the E. coli DXS system to orthologue(s) fromplant species that will be more relevant model systems for herbicidal inhibitordevelopment. In addition to NMR, we will use structural MS (HDX-MS, fast photochemicaloxidation of proteins (FPOP)-MS and ion-mobility spectrometry- (IMS)-MS to map the binding sites and modes of interaction of substrates, catalytic / transition state intermediates and inhibitors. Molecular modelling based on existing PDB structures and AlphaFold2 structural models will be used as a basis for incorporating our experimentally-derived restraints.

在利兹大学和先正达PLC之间的这4年的BBSRC ICase博士学位中，该学生将研究对农业知识行业重要的植物酶的结构，功能和机制。该项目由Alex Breeze和Frank Sobott教授和弗兰克·索博特（Alex Breeze）和弗兰克·索博特（Frank Sobott）共同监督在艾斯伯里结构分子生物学中心，基于正弦和利兹大学和利兹大学之间的先前合作，进一步阐明了机械生物学和抑制型tpp path tpp-tppe依赖性的抑制模式。合酶（DXS）。结果将是对潜在抑制剂设计策略和机制的增强理解，这可能会导致对针对这一途径的改善的改良的草本化剂的发展。尽管属于一系列普遍理解的酶，但对DXS的机械理解，对DXS的机械理解及其抑制作用仍然相对限制，因为在结构上，尤其是结构性的确定，或者是结构性的，或者是结构性的，或者是核对的，或者是核对的，或者是核对的，或者是核对的，或者是核对的，或者是态度的，或者是核对的，或者是核对的。事实证明，X射线晶体学非常具有挑战性。该学生将使用尖端的超高野外甲基NMR和氢 - 甲基升华质谱法（HDX-MS）来建立我们的早期工作。具体而言，我们将基于大肠杆菌DXS系统的甲基NMR方法转移到植物物种的直系同源物，这些物种将成为植物抑制层的更相关的模型系统。除NMR外，我们还将使用结构性MS（HDX-MS，蛋白质（FPOP）-MS的快速光化学氧化和离子动力谱图 - （IMS）-MS来映射基质的结合位点和相互作用的结合位点和模式。纳入我们的实验衍生的约束的基础。