Probing enzyme reaction cascades in polyketide biosynthesis at atomic resolution by NMR

通过 NMR 在原子分辨率下探测聚酮化合物生物合成中的酶反应级联

• 批准号: 2770488
• 金额: --
• 依托单位: University of Bristol
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2770488
• 关键词: Probing; enzyme; reaction; cascades; polyketide

Many antibiotics and other potentially therapeutic products are produced by type I polyketide synthases. These systems work like assembly lines in order to build up molecules, such that by manipulating enzymes within the cascade, the end product can be altered. By advancing our understanding of these enzymes, we can even begin to look at using them for the design of novel molecules.One system of particular interest for this project is the bongkrekic acid system, named after the toxin it produces. Bongkrekic acid works by inhibiting the adenine nucleotide translocase (ANT), blocking oxidative phosphorylation. Interestingly, compounds which selectively target the ANT have been put forward as antiapoptotic agents that may be used in chemotherapy. Bongkrekic acid therefore represents an excellent starting structure for this rational development, a process which this work will provide key information for. It will highlight how synthesis and flux is controlled by the enzymes, and therefore how this may be manipulated to produce analogues. Key questions in the project surround how the enzymes control which beta branching events include a final decarboxylation and how synthesis is terminated without the presence of a terminal thioesterase enzyme. Novel NMR assays and supplementary ESI-MS will be used that allow monitoring of intermediates in the enzyme cascade. This allows probing of bongkrekic acid synthesis at atomic resolution, without the need for these intermediates to be individually isolated. Where appropriate, structural techniques for analysis of protein interactions may be used, such as x-ray crystallography and cryo-EM. This project falls within the EPSRC Chemical Biology and Biological Chemistry research area.

许多抗生素和其他潜在的治疗产物都是由I型聚酮化合物合成酶产生的。这些系统像组装线一样工作，以建立分子，以便通过操纵级联体内的酶，可以改变最终产物。通过促进我们对这些酶的理解，我们甚至可以开始考虑使用它们来设计新分子。该项目特别感兴趣的系统是以其产生的毒素命名的Bongkrekic Acid系统。 Bongkrekic Acid通过抑制腺嘌呤核苷酸易位酶（ANT）的作用，阻断氧化磷酸化。有趣的是，选择性靶向蚂蚁的化合物已被提交为可以用于化学疗法的抗凋亡剂。因此，Bongkrekic Acid代表了这种合理发展的出色启动结构，这项工作将为这项工作提供关键信息。它将强调合成和通量如何由酶控制，因此如何操纵它以产生类似物。该项目中的关键问题围绕着酶如何控制哪种β分支事件包括最终的脱羧以及如何在没有末端硫酯酶酶的情况下终止合成。将使用新型的NMR分析和补充ESI-MS，以监测酶级联中的中间体。这允许在原子分辨率下探测bongkrekic酸合成，而无需单独分离这些中间体。在适当的情况下，可以使用用于分析蛋白质相互作用的结构技术，例如X射线晶体学和冷冻EM。该项目属于EPSRC化学生物学和生物化学研究领域。