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品是由I型聚酮合成酶生产的。为了构建分子，这些系统就像装配线一样工作，这样通过操纵级联中的酶，最终产品就可以改变。通过加深我们对这些酶的了解，我们甚至可以开始考虑用它们来设计新的分子。这个项目特别感兴趣的一个系统是bongkrekic酸系统，以它产生的毒素命名。Bongkrekic酸通过抑制腺嘌呤核苷酸转位酶（ANT），阻断氧化磷酸化起作用。有趣的是，选择性靶向ANT的化合物已被提出作为抗凋亡药物，可能用于化疗。因此，Bongkrekic酸代表了这种合理发展的良好起始结构，这项工作将为这一过程提供关键信息。它将强调如何合成和通量是由酶控制的，因此如何操纵它来产生类似物。该项目的关键问题围绕酶如何控制哪些β分支事件包括最终脱羧以及如何在没有末端硫酯酶存在的情况下终止合成。新的核磁共振分析和补充ESI-MS将用于监测酶级联中的中间体。这使得在原子分辨率下探测bongkrekic酸合成，而不需要单独分离这些中间体。在适当的情况下，可以使用结构技术来分析蛋白质相互作用，如x射线晶体学和低温电镜。该项目属于EPSRC化学生物学和生物化学研究领域。