Determining the reductive pathway in the endoplasmic reticulum of mammalian cells

细胞正确折叠和组装蛋白质的能力是蛋白质合成的最后阶段。蛋白质折叠需要一组能够催化折叠反应或充当分子伴侣的蛋白质，以防止非生产性蛋白质聚集和细胞应激。细胞无法进行折叠过程会导致细胞死亡，从而导致一些最灾难性的疾病病理，如糖尿病、阿尔茨海默氏症和帕金森氏症。为了保持细胞和组织的健康，它们必须能够制造蛋白质，并且它们制造的蛋白质必须能够正确发挥作用。细胞拥有复杂的机制，以确保当新的蛋白质产生时，它们具有功能，并被运输到正确的位置，无论是在细胞内还是细胞外。这个项目将确定一个关键的过程，使蛋白质能够有效地制造并被输送到细胞外，特别是这个过程在疾病期间如何分解。蛋白质的生产和输送可以概括为两个关键阶段：i)确保蛋白质的正确制造和采用正确的形状，ii)蛋白质从细胞内到细胞外的运输。蛋白质是由一串氨基酸组成的，这些氨基酸卷曲或折叠形成具有特征的形状或三维结构。只有一个这样的形状是有功能的，细胞通过提供辅助蛋白或伴侣来帮助这一过程来确保采用这种形状。如果细胞不能正确折叠蛋白质，就会导致疾病。为了让分泌的蛋白质发挥作用，它们需要强大，并确保它们在蛋白质内形成连接，将蛋白质连接在一起。这些键被称为二硫键。如果没有这些键，这些蛋白质就不会发挥作用，也不会分泌。我们对如何引入正确的链接或如何删除不正确的链接知之甚少。这个项目将研究正确的键是如何形成的。我们的小组想要详细了解细胞如何提供正确的环境来允许蛋白质折叠并形成正确的二硫键以确保它们的稳定性。为了了解细胞如何折叠和组装蛋白质，我们使用细胞生物学和生化技术的组合来研究哺乳动物细胞的这一过程。

The ability of cells to correctly fold and assemble proteins is the final stage in protein synthesis. Protein folding requires a subset of proteins able to either catalyse folding reactions or act as molecular chaperones preventing non-productive protein aggregation and cell stress. The inability of cells to carry out the folding process results in cell death and consequently some of the most catastrophic disease pathologies such as diabetes, Alzheimer's and Parkinson's.For cells and tissues to remain healthy they must be able to make proteins and the proteins they make must be able to function correctly. The cell has complex machinery for ensuring that when new proteins are made they are functional and are transported to the correct location, be it within the cell or outside. This project will determine one crucial process that allows proteins to be made efficiently and be delivered outside the cell and, in particular, how this process breaks down during disease. The production and delivery of proteins can be summarised into two key stages: i) ensuring proteins are made correctly and adopt the correct shape, ii) transport of the proteins from the inside to the outside of the cell.Proteins are made as a string of amino acids which coil-up or fold to adopt a characteristic shape or three-dimensional structure. Only one such shape is functional and the cell ensures that this shape is adopted by providing helper proteins or chaperones to aid this process. If cells are unable to correctly fold proteins then disease results. For the secreted proteins to function they need to be robust and to ensure this is the case they form links within the protein to tie the protein together. These links are called disulfide bonds. Without these bonds the proteins would not function and would not be secreted. We know very little about how the correct links are introduced or how incorrect linkages are removed. This project will investigate how the correct bonds are formed. Our group wants to understand in detail how cells provide the correct environment to allow proteins to fold and to form the correct disulfide bonds to ensure their stability. To understand how cells fold and assemble proteins we are studying this process in mammalian cells using a combination of cell biological and biochemical techniques.