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

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许多重要的应用。它们被用来对付威胁生命的疾病，如细菌和真菌感染、器官移植排斥和癌症，并作为除草剂、杀虫剂和杀真菌剂，在保护粮食作物方面发挥着重要作用。许多天然产品是由酶“装配线”组装的，类似于汽车生产线。装配线的每个组件都必须与下一个组件进行有效的沟通，以确保整个流程的效率。这种通信通常由附接到组件末端的专用“对接域”介导，所述组件以特定方式彼此相互作用。我们一直在研究一种特定类型的对接结构域和它的相互作用伴侣在enacloxin IIa的组装中所发挥的作用。这种抗生素由伯克霍尔德菌属产生，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治疗方法。我们的数据表明，这种类型的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，包括临床上使用的几种抗癌药物和抗生素。在这个项目中，我们的目标是使用已建立和最近开发的技术相结合，以更好地了解参与enacloxin生产的对接结构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enacyloxin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的系统，其中两个组件，这两个组件都有一个对接域，这是一个类似的参与enacloxin生产，似乎与相同的合作伙伴进行交互，执行顺序的任务，在装配的aerodynosins，一个不寻常的组蛋白质降解抑制剂产生的蓝藻。这将拓宽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相互作用伙伴的方式的共同原则。最后，我们将研究是否可以利用我们对这些共同原理的理解，用来自enacloxin系统的相应组分替代aerodosin组装线的组分之一。总的来说，这个项目将显着加深我们的理解所发挥的作用的一个重要类型的对接域在天然产物组装，并建立利用这种对接域构建工程装配线能够生产新的天然产物杂交的可行性。

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.