Mapping Protein-Protein Interactions in Modular Polyketide Synthases by Carbene Footprinting

来自植物和微生物的生物活性天然产物在医学和农业中具有不可或缺的应用。它们通常用于治疗危及生命的疾病，包括细菌、真菌和病毒感染以及癌症。此外，这些有价值的分子还可用作除草剂、杀虫剂和杀真菌剂，对保护粮食作物至关重要。这些化合物中的许多是由巨大的酶“装配线”组装的，就像汽车生产线一样，化合物以合乎逻辑的逐步方式构建。在生产过程中，装配线的每个组件都必须与装配线上的其他酶进行生产性沟通。这些相互作用对于确保整个过程的有效性和保持产品保真度至关重要。在这个项目中，我的目标是使用已建立和最近开发的技术相结合，以更好地了解驱动这些生物合成过程的相互作用。我的重点将涉及生产一种名为quartomicin的强效抗病毒化合物的酶，quartomicin靶向病毒逆转录酶，是一个更大的化合物家族的一部分，称为tetronates。这个特殊的系统提供了一个机会，研究一系列的相互作用，重要的生产含特膦酸盐的天然产物。这将拓宽我们对蛋白质-蛋白质相互作用在天然产物组装中所起作用的理解，并使我们能够建立它们识别其相互作用伙伴的方式的共同原则。这项研究的结果将揭示可用于生产更有效的生物催化酶的共同原理，并通过扩展生物活性聚酮化合物的新型衍生物。这个项目将大大加深我们对蛋白质所起作用的理解，蛋白质的相互作用，并将建立合理的基础，利用这种相互作用，以构建工程组装线，能够产生新的天然产物类似物。

Bioactive natural products from plants and microorganisms have indispensable applications in both medicine and agriculture. They are often employed to treat life-threating conditions, including bacterial, fungal and viral infections and cancer. Moreover, these valuable molecules are also used as herbicides, insecticides and fungicides that are essential for the protection of food crops. Many of these compounds are assembled by giant enzymatic 'assembly lines', much like a car production line, with the compound being constructed in a logical stepwise manner. During production, each component of the assembly line must engage in productive communication with other enzymes in the assembly line. These interactions are critical to ensuring that the overall process is efficient and maintains product fidelity. In this project, I aim to use a combination of established and recently-developed techniques to develop a better understanding of the interactions driving these biosynthetic processes. My focus will concern the enzymes involved in production of a potent antiviral compound called quartomicin, which targets the viral reverse transcriptase and is part of a larger family of compounds called tetronates. This particular system provides the opportunity to study an array of interactions important for producing tetronate-containing natural products. This will broaden our understanding of the role played by protein-protein interactions in natural product assembly and allow us to establish the common principles underlying the way in which they recognise their interaction partners. The results of this research will reveal common principles that can be harnessed to produce more efficient enzymes for biocatalysis, and by extension novel derivatives of bioactive polyketides.Overall, this project will significantly deepen our understanding of the roles played by protein-protein interactions during natural product assembly and will establish a rational basis for exploiting such interactions to construct engineered assembly lines capable of producing novel natural product analogues.