Dissecting and Harnessing Carrier Protein Interactions in Fungal Megasynth(et)ases

解析和利用真菌大合成酶中载体蛋白的相互作用

• 批准号: MR/W011247/1
• 负责人: Matthew Jenner
• 金额: $ 162.11万
• 依托单位: University of Warwick
• 依托单位国家: 英国
• 项目类别: Fellowship
• 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=MR%2FW011247%2F1
• 关键词: Dissecting; Harnessing; Carrier; Protein; Interactions

Fungal polyketide synthases (PKS) are giant multi-domain proteins responsible for the biosynthesis of a vast number of biologically active natural products. Whilst many find important applications as medicinal or agrochemical agents, some are toxins that are harmful to both human health and agriculture. The precise assembly of these complex compounds relies on the highly programmed protein-protein interactions (PPIs) between a small carrier protein (CP) domain, to which biosynthetic intermediates are covalently tethered, and the individual catalytic domains within the PKSs, ensuring that the overall process is efficient and maintains product fidelity. Despite the importance of these interactions, they remain poorly understood, primarily due to their ephemeral nature, requiring a combination of techniques to be studied effectively. Uncovering the molecular factors governing programming is the greatest remaining problem in our understanding of fungal PKSs, and represents a huge obstacle to rewiring these enzymes towards user-designed molecules. This project aims to combine cutting-edge mass spectrometry, structural biology and biochemical techniques to elucidate the molecular details of CP-dependent interactions underpinning fungal PKS machinery. The research will initially focus on characterising PPIs in two similar PKS systems; one involved in the biosynthesis of cyclosporin (clinically used immunosuppressant), the other responsible for the construction of lovastatin (clinically used cholesterol-lowering agent). These PKSs differ by the positioning of a single catalytic domain, and therefore represent excellent model systems to establish the common principles underlying CP-dependent interactions. Taken together, this body of work will significantly deepen our understanding of the roles played by PPIs in fungal PKSs. It will also form a knowledge-base to begin exploiting such interactions to construct engineered systems capable of producing novel natural product analogues.

真菌聚酮酶（PKS）是一种多结构域的蛋白质，参与了大量具有生物活性的天然产物的生物合成。虽然许多被发现作为医药或农业化学剂的重要应用，但有些是对人类健康和农业有害的毒素。这些复杂化合物的精确组装依赖于小载体蛋白（CP）结构域（生物合成中间体与其共价连接）和PKS内的单个催化结构域之间的高度程序化蛋白质-蛋白质相互作用（PPI），确保整个过程有效并保持产品保真度。尽管这些相互作用的重要性，他们仍然知之甚少，主要是由于其短暂的性质，需要一个有效的研究技术相结合。揭示控制编程的分子因素是我们理解真菌PKS的最大剩余问题，并且代表了将这些酶重新连接到用户设计的分子的巨大障碍。该项目旨在结合联合收割机尖端的质谱，结构生物学和生物化学技术，阐明CP依赖的相互作用的分子细节支撑真菌PKS机制。该研究最初将集中在两个类似PKS系统中的PPI特征;一个参与环孢菌素（临床使用的免疫抑制剂）的生物合成，另一个负责洛伐他汀（临床使用的降胆固醇剂）的构建。这些PKS不同的定位一个单一的催化结构域，因此代表了优秀的模型系统，以建立CP依赖的相互作用的共同原则。两者合计，这一机构的工作将显着加深我们对PPIs在真菌PKSs中所发挥的作用的理解。它还将形成一个知识基础，开始利用这种相互作用来构建能够产生新的天然产物类似物的工程系统。

项目成果

Elucidating the molecular programming of a nonlinear non-ribosomal peptide synthetase responsible for fungal siderophore biosynthesis.

• DOI: 10.1038/s41467-023-38484-8
• 发表时间: 2023-05-17
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Jenner, Matthew;Hai, Yang;Nguyen, Hong H.;Passmore, Munro;Skyrud, Will;Kim, Junyong;Garg, Neil K.;Zhang, Wenjun;Loo, Rachel R. Ogorzalek R.;Tang, Yi
• 通讯作者: Tang, Yi