Deciphering the conformational mechanisms of nascent membrane protein folding

细胞膜是主要由蛋白质和脂质组成的动态结构，其作用是将细胞区室化，为细胞及其细胞器的外部环境提供屏障。整合膜蛋白存在于细胞膜内，并负责各种动态过程，如感觉、细胞调节和细胞间粘附。膜蛋白的功能能力和它们的表达水平将在很大程度上决定离子组成，从而决定给定细胞类型的代谢水平，使它们对所有生命都是必不可少的。膜蛋白必须被“正确”折叠才能发挥功能。这项研究的主要目的是了解新生膜蛋白如何折叠。人们对膜蛋白在产生过程中和细胞膜背景下折叠的化学原理知之甚少。研究膜蛋白是一项艰巨的任务，由于膜蛋白的顽固性和疏水性，相比之下，其可溶性蛋白质的对应部分;与额外的蛋白质机器被要求为他们的细胞定位和折叠内的膜，相比之下。了解所涉及的过程将增强我们对细胞如何获得“正确”折叠和功能性膜蛋白的理解，并假设“错误”折叠和聚集的膜蛋白是如何形成的-这是一个对细胞有毒的过程。因此，开发反映细胞过程的稳健测定将使辅因子、突变和药物对膜蛋白折叠过程的影响成为可能。此外，还有一个关键目标是开发方法，以获得结构洞察膜蛋白事件使用先进的化学工具和技术，如氢-氘交换质谱。这项研究拟在伦敦国王学院，化学系，保拉布斯教授的实验室内进行。该研究将涉及生产能够在体外生产，插入和折叠膜蛋白的系统，并开发这些过程的结构评估方法，重点是使用氢氘交换质谱法。

Cellular membranes are dynamic structures consisting mostly of protein and lipid which act to compartmentalise the cell, providing barriers to the external environments of the cell and its organelles. Integral membrane proteins reside within a cellular membrane and are responsible for a variety of dynamic processes, such as sensation, cellular regulation, and cell-to-cell adhesion. A membrane protein's functional capability and their level of expression will largely decide the ionic composition, and therefore the metabolic levels of a given cell type, making them essential for all life. Membrane proteins must be folded 'correctly' to become functional. The key aim of the research proposal is to understand how nascent membrane proteins fold. The chemical principles of membrane protein folding during their production and in the context of a cellular membrane are poorly understood. Investigating membrane proteins is a difficult task due to the intractable and hydrophobic nature of membrane proteins, compared to their soluble protein counter parts; with additional protein machinery being required for their cellular localisation and folding within the membrane, in comparison. To understand the processes involved would enhance our understanding on how the cell obtains 'correctly' folded and functional membrane proteins, and postulate on how 'incorrectly' misfolded and aggregated membrane proteins are formed - a toxic process for a cell. Consequently, development of robust assays that reflect the cellular processes will enable the influence of co-factors, mutations and drugs on the process of membrane protein folding. Moreover, there is a key aim to develop methodologies to gain structural insight into membrane protein events using advanced chemical tools and techniques, such as hydrogen-deuterium exchange mass spectrometry.The research is proposed to take place at the King's College London, Department of Chemistry, within the laboratory of Professor Paula Booth. The research will involve producing systems capable of both membrane protein production, insertion and folding in vitro and developing methodologies for the structural assessment of these processes with strong focus on using hydrogen deuterium exchange mass spectrometry.