Structure and function of key ABC Transporters in health and disease

人ABC转运蛋白是52个完整的膜蛋白家族，可跨细胞膜运输不同的分子。ABC转运蛋白的突变导致一系列疾病，包括糖尿病(ABCC8和ABCC9)、囊性纤维化(ABCC7、CFTR)和失明(ABCA4)。Liz Carpenter和Heidi de Wet的团队研究与疾病有关的人类ABC转运蛋白的3D结构和功能。特别是，利兹·卡彭特的团队研究了一系列与神经精神疾病、癌症、罕见和代谢性疾病以及炎症性疾病有关的蛋白质。尽管膜蛋白的研究被认为是具有挑战性的，但这一领域刚刚随着新的技术发展而打开大门，包括生产蛋白质的新方法，用于X射线结晶学的结晶方法，以及通过电子冷冻显微镜获得高分辨率结构的方法。由于人类完整的膜蛋白在蛋白质水平上的研究具有挑战性，学生将从一系列项目开始，开发2到3个难度不同的项目，然后选择最感兴趣和表现良好的蛋白质进行研究。我们确实有一些已经纯化了蛋白质的项目，以及其他有趣的目标，这些项目需要在这方面进行开发。这个项目将涉及生产一系列人类ABC转运蛋白，特别是那些与疾病相关的转运蛋白。然后，学生将使用冷冻-EM技术研究它们的结构，其中包括一系列核苷酸、核苷酸类似物、底物和抑制剂。这些结构将提供对这些蛋白质如何发挥作用的原子水平的了解。他们还将使用功能分析，包括ATPase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，他们在生产和研究复杂蛋白质的结构方面拥有专业知识。他们也是开发化验的专家，以确定小分子稳定剂和转运蛋白的激活剂，这是学生将使用的技术。学生将获得ABC转运体结构和功能研究的丰富应用知识，包括生化(ATPase活性、转运体分析)和生物物理方法(热稳定性、微尺度热电泳法，SPR)以及结构技术，包括冷冻-EM和X射线结晶学。这名学生将花大部分时间在牛津SGC的Liz Carpenter教授的团队中，与14名膜蛋白结构生物学家组成的团队一起工作。他们还将与牛津DPAG的Heidi de Wet教授合作进行功能研究。该项目提供了一个与热心的行业合作伙伴广泛合作的绝佳机会，包括与您在美国波士顿的行业主管一起工作的3到4个月的访问。

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.