Elucidating and exploiting docking domain-mediated carrier protein recognition in natural product megasynthetases

来自植物和微生物的具有生物活性的天然产物在医药和农业中有许多重要的应用。它们被用于应对威胁生命的疾病，如细菌和真菌感染、器官移植排斥反应和癌症，并被用作除草剂、杀虫剂和杀菌剂，在保护粮食作物方面发挥着重要作用。许多天然产品是通过酶的“装配线”组装的，类似于汽车生产线。装配线上的每个组件都必须与下一个组件进行有效的沟通，以确保整个过程的效率。这种通信通常由连接到组件末端的专用“对接域”介导，它们以特定的方式相互作用。我们一直在研究一种特殊类型的对接结构域及其相互作用伙伴在乙酰胆碱IIa的组装中所起的作用。这种抗生素由伯克霍尔德菌产生，对鲍曼不动杆菌具有强效活性，鲍曼不动杆菌是一种导致人类生命威胁疾病的细菌，迫切需要找到有效的新疗法。我们的数据表明，这种对接结构域参与了比以前认为的更多的生物活性天然产物的组装，包括几种抗癌药物和临床使用的抗生素。在这个项目中，我们的目标是结合现有的和最近开发的技术来更好地理解参与生产的对接域识别其相互作用伙伴的方式。我们获得的见解将用于修改乙酰胆碱装配线，看看它是否按照我们预测的方式运行。我们还旨在研究一个相关系统，其中两个组件，这两个组件都有一个对接域，类似于一个参与制定的制定，似乎与相同的伙伴相互作用，以执行顺序的任务，在组装铜绿蛋白苷，一组不寻常的蛋白质降解抑制剂由蓝藻产生。这将扩大我们对对接域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交互伙伴方式的共同原则。最后，我们将研究我们对这些共同原理的理解是否可以利用绿脓毒杆菌素装配线的一个组件来替代来自乙酰胆碱素系统的相应组件。总体而言，该项目将显著加深我们对一种重要对接域在天然产物组装中所起作用的理解，并将建立利用这种对接域构建能够生产新型天然产物杂交的工程装配线的可行性。

Bioactive natural products from plants and microorganisms have numerous important applications in medicine and agriculture. They are used to tackle life-threating conditions, such as bacterial and fungal infections, organ transplant rejection and cancer, and as herbicides, insecticides and fungicides that play an essential role in the protection of food crops. Many natural products are assembled by enzymatic "assembly lines", akin to a car production line. Each component of the assembly line must engage in effective communication with the next to ensure the overall process is efficient. Such communication is typically mediated by dedicated "docking domains" attached to the ends of the components, which interact with each other in a specific way. We have been studying the role played by a particular type of docking domain and its interaction partner in the assembly of enacyloxin IIa. This antibiotic is produced by Burkholderia species and has potent activity against Acinetobacter baumannii, a bacterium that causes life threatening diseases in humans for which there is a critical need to find effective new treatments. Our data have shown that this type of docking domain is involved in the assembly of many more bioactive natural products than previously thought, including several anti-cancer agents and an antibiotic that are used in the clinic. In this project we aim to use a combination of established and recently-developed techniques to develop a better understanding of the way the docking domain involved in the production of enacyloxin recognises its interaction partner. The insights we obtain will be used to modify the enacyloxin assembly line to see whether it behaves in the way we predict. We also aim to investigate a related system in which two components, both of which have a docking domain that is similar to the one involved in enacyloxin production, appear to interact with the same partner to execute sequential tasks in the assembly of the aeruginosins, an unusual group of protein degradation inhibitors produced by cyanobacteria. This will broaden our understanding of the role played by docking domains in natural product assembly and allow us to establish the common principles underlying the way in which they recognise their interaction partners. Finally, we will investigate whether our understanding of these common principles can be harnessed to substitute one of the components of the aeruginosin assembly line with the corresponding component from the enacyloxin system. Overall, this project will significantly deepen our understanding of the roles played by an important type of docking domain in natural product assembly and will establish the feasibility of exploiting such docking domains to construct engineered assembly lines capable of producing novel natural product hybrids.