Harnessing Nature's ability to create membrane compartmentalisation through redesign of a protein machinery.

生物细胞可以被认为是一个复杂的化学反应器，其中大量的相互作用同时发生;这些对它的功能和生存至关重要。为了防止所有这些并发化学途径的“噪音”中不必要的串扰和干扰，细胞将这些过程划分为各个膜结合结构（细胞器）中的不同功能。化学过程的限制也允许细胞维持不相容的环境，这些环境对每个细胞器的功能都是最佳的，这在单个“锅”中是不可能的。如果我们要在合成“纳米反应器”中模拟这种复杂性，甚至构建能够在医学，环境修复或能量捕获和存储等不同领域执行有用任务的合成细胞，我们将需要开发在合成人造结构中模拟细胞区室化的方法。这是我们在这个项目中将要面对的挑战。活细胞内的内部膜结合区室通常是由“蛋白质”成分组成的分子机器产生的。我们将重新设计属于这些类型的机器之一的蛋白质的结构，以控制其与实验室中制造的膜囊泡的可用性。最终，我们将使用这种基于蛋白质的生物技术来创建包含在更大的膜容器（细胞器）中的纳米反应器。在高等生物（如人类）中自然发生的膜成形机制由许多蛋白质协同工作组成;显然，这种复杂性目前与大规模实验室工程的分隔结构不兼容。因此，我们项目的第一个目标是设计一个最小的膜成形蛋白系统，作用于高等生物膜。这种生物简单性的先例已经在一些古老的细菌中观察到，这些细菌含有由非常少的蛋白质成分组成的类似的膜成形机器。我们将通过基因工程减少一个已经很好表征的高等生物复合体的功能成分的数量来模仿这个更简单的模型。这种基于蛋白质的技术将用于开发类似于生物细胞的多隔室结构，每个隔室包含不同大小和化学性质的分子。我们将学会控制这种生物技术的膜成形作用，以便我们能够停止和重新开始新隔室的产生。这将使我们能够控制不同成分在更大的膜结构内的特定隔室中的封装。第一个概念验证将包括封装一种能够将遗传信息转换为新蛋白质分子的分子机器，在一个单一的内部隔室中，模仿细胞核的功能。将开展进一步的工作，以允许化学品在单个容器内的内部隔室之间选择性地移动。该项目的最终产品将是一个功能齐全的工具包，使研究人员能够创建复杂的分区和通信纳米反应器。这些装置可以用作细胞大小的载体，同时产生，运输和递送治疗货物到体内的特定靶点。或者，它们可以被用作化学工厂，可以从供水中去除污染物。

A biological cell can be thought of as a complex chemical reactor where vast numbers of interactions are simultaneously taking place; these are vital to its function and survival. To prevent unwanted cross-talk and interference within the "noise" of all these concurrent chemical pathways, a cell compartmentalises these processes localizing different functions within individual membrane-bound structures (organelles). Confinement of chemical processes also allows a cell to maintain incompatible environments that are optimal for each organelle's function, which would not be possible within a single "pot".If we are to mimic this complexity within synthetic "nanoreactors", or even build synthetic cells capable of useful tasks within areas as diverse as medicine, environmental remediation, or energy capture and storage, we will need to develop ways of mimicking cellular compartmentalisation within synthetic man-made structures. This is the challenge we will address in this project.Internal membrane bound compartments within living cells are normally generated by molecular machineries made up of 'protein' components. We will redesign the structure of proteins belonging to one of these types of machineries to control its usability with membrane vesicles made in the laboratory. Ultimately, we will use this protein-based biotechnology to create nanoreactors contained within the larger membrane vessel (organelle). Membrane-shaping machineries naturally occurring in higher organisms, such as humans, are composed of many proteins working in concert; obviously such complexity is not currently compatible with large scale laboratory engineering of compartmentalised architectures. Therefore the first aim of our project is to engineer a minimal membrane-shaping protein system working on higher organism membranes. Precedent for this biological simplicity has been observed in some ancient bacteria containing similar membrane-shaping machineries made up of very few protein components. We will mimic this simpler model by reducing the number of functional components of an already well-characterised higher organism complex, through genetic engineering. This protein-based technology will be used to develop multi-compartment architectures, similar to biological cells, with each compartment containing molecules of different sizes and chemical properties. We will learn to control the membrane shaping action of this biotechnology so that we are able to stop and restart the generation of new compartments. This will allow us to control the encapsulation of different components in specified compartments within the larger membrane structure.A first proof of concept will consist of encapsulating a molecular machinery capable of converting genetic information into new protein molecules inside one single internal compartment, mimicking the function of the cell nucleus. Further work will be conducted to allow selective movement of chemicals between internal compartments within a single vessel. The ultimate product of this project will be a well-characterised, functional toolkit enabling researchers to create complex compartmentalised and communicating nanoreactors. These devices could be used as cell-sized vehicles that simultaneously produce, transport and deliver therapeutic cargo to specific targets in the body. Alternatively, they could be employed as chemical factories that can remove pollutants from water supplies.