Structure and function of key ABC Transporters in health and disease

健康和疾病中关键 ABC 转运蛋白的结构和功能

• 批准号: 2270392
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2270392
• 关键词: Structure; function; key; ABC; Transporters

Human ABC transporters are a family 52 integral membrane proteins that transport diverse molecules across cell membranes. Mutations in ABC transporters causes a range of diseases, including diabetes (ABCC8 and ABCC9), cystic fibrosis (ABCC7, CFTR) and blindness (ABCA4).At the SGC in Oxford Profs. Liz Carpenter and Heidi de Wet's groups study the 3D structures and function of human ABC transporters, involved in diseases. In particular Liz Carpenter's group works with a range of proteins involved in neuropsychiatric disease, cancer, rare and metabolic disease, as well as inflammatory diseases. Although membrane proteins are thought to be challenging to study, this field is just now opening up with new technical developments, including novel methods for producing proteins, for crystallisation for X-ray crystallography and for obtaining high resolution structures by electron cryo-microscopy.As human integral membrane proteins are challenging to study at the protein level, the student would start with a range of projects, develop 2 or 3 projects of different levels of difficulty and then select the most interesting and well-behaved proteins to work on. We do have a number of projects where the proteins are already purified, as well as other interesting targets where this aspect of the project would require development.This project would involve production of a range of human ABC transporters, particularly those associated with disease. The student would then study their structures using cryo-EM with a range of nucleotides, nucleotide analogues, substrates and inhibitors. These structures will provide an atomic level understanding of how these proteins function. They would also use functional assays, including ATPase activity assays, transport assays and biophysical assays to further their understanding of the function of these proteins in health and disease. This work is in collaboration with supervisors at a major pharmaceutical company, who have expertise in producing and studying the structures of complex proteins. They are also experts in developing assays to identify small molecule stabilizers and activators for transporters, techniques that would be used by the student. The student will gain a strong working knowledge of ABC transporter structural and functional studies, including biochemical (ATPase activity, transport assays) and biophysical methods (thermostability, microscale thermophoresis, SPR) and structural techniques, including cryo-EM and X-ray crystallography. The student will spend most of their time working in Prof. Liz Carpenter's group at the SGC, in Oxford, with a team of 14 membrane protein structural biologists. They will also collaborate with Prof. Heidi de Wet, in DPAG, Oxford, for functional studies. This project provides an outstanding opportunity to work extensively with enthusiastic collaborators in industry, including a 3 or 4 months visit to work with your industrial supervisors in Boston, USA.

人ABC转运蛋白是一个家族，52个完整的膜蛋白，跨细胞膜转运不同的分子。ABC转运蛋白的突变会导致一系列疾病，包括糖尿病（ABCC 8和ABCC 9）、囊性纤维化（ABCC 7、CFTR）和失明（ABCA 4）。Liz Carpenter和Heidi de Wet的团队研究了与疾病有关的人类ABC转运蛋白的3D结构和功能。特别是Liz Carpenter的团队研究了一系列涉及神经精神疾病，癌症，罕见和代谢疾病以及炎症性疾病的蛋白质。虽然膜蛋白被认为是具有挑战性的研究，这一领域是刚刚开放与新的技术发展，包括新的方法来生产蛋白质，结晶的X射线晶体学和获得高分辨率的结构，通过电子冷冻显微镜。由于人类的整体膜蛋白是具有挑战性的研究在蛋白质水平上，学生将开始一系列的项目，开发2到3个不同难度的项目，然后选择最感兴趣和表现最好的蛋白质进行研究。我们确实有一些已经纯化蛋白质的项目，以及其他需要开发的项目。这个项目将涉及一系列人类ABC转运蛋白的生产，特别是那些与疾病相关的转运蛋白。然后，学生将使用cryo-EM与一系列核苷酸，核苷酸类似物，底物和抑制剂一起研究它们的结构。这些结构将提供对这些蛋白质功能的原子水平理解。他们还将使用功能分析，包括ATP酶活性分析，运输分析和生物物理分析，以进一步了解这些蛋白质在健康和疾病中的功能。这项工作是与一家大型制药公司的主管合作进行的，他们拥有生产和研究复杂蛋白质结构的专业知识。他们也是开发鉴定转运蛋白小分子稳定剂和激活剂的检测方法的专家，这些技术将由学生使用。学生将获得ABC转运蛋白结构和功能研究的强大工作知识，包括生物化学（ATP酶活性，运输测定）和生物物理方法（热稳定性，微尺度热泳，SPR）和结构技术，包括cryo-EM和X射线晶体学。该学生将花大部分时间在牛津大学SGC的Liz Carpenter教授的团队中工作，该团队由14名膜蛋白结构生物学家组成。他们还将与牛津大学DPAG的Heidi de Wet教授合作进行功能研究。该项目提供了一个与行业中热情的合作者广泛合作的绝佳机会，包括3或4个月的访问，与您在美国波士顿的工业主管一起工作。