The role of adaptor proteins in the dynamics of clathrin coat formation and disassembly

课程概述：这个由MRC资助的博士培训伙伴关系(DTP)将尖端分子和分析科学与数据分析中的创新计算方法结合在一起，使学生能够解决假设引导的生物医学研究问题。这是一个为期4年的课程，第一年包括一系列的教学模块和两个基于实验室的研究项目，这些项目导致了跨学科生物医学研究的硕士学位。前两个学期包括精选的教学模块，使学生能够在多学科的科学中获得坚实的基础。学生们还参加了由学术和行业专家主持的一系列大师课程，这些专家涉及分子、细胞和组织动力学、微生物学和传染病、应用生物医学技术以及人工智能和数据科学等领域。在第三学期和暑期，学生们在他们选择的实验室里进行两个为期十一周的研究项目。项目：笼蛋白介导的内吞作用是分子选择性进入细胞的主要途径之一，在突触小泡再循环、受体再循环、信号传递和发育等广泛的基本过程中发挥核心作用。这是通过形成一层蛋白质状的网状蛋白涂层来实现的，该涂层由网状蛋白与接头蛋白的相互作用控制，导致囊泡的萌发和形成。虽然我们对单个接头蛋白在内吞作用中的作用的了解越来越多，但我们仍然不清楚多个接头蛋白与笼状蛋白相互作用的方式，也不了解它们相互作用的分子细节。主要的两个目标是：1.利用动态光散射、荧光光谱和表面等离子体共振等多种生物物理技术，研究关键接头蛋白如何影响组装和拆卸的动力学。使用高分辨率低温电子显微镜和单粒子分析技术对连接到适配器上的笼子进行三维结构分析，以可视化关键适配器与组装的笼子的结合模式。在实习期间，将培养来自不同学科的几项技能，包括如何进行表面等离子体共振、动态光散射和荧光光谱等生物物理技术，并学习如何分析从这些生物物理技术获得的动力学数据。开发的结构和分子生物学技术将包括蛋白质纯化和表达、蛋白质凝胶分析(SDS-PAGE)、GST下拉、高分辨率冷冻电子显微镜和单颗粒分析。这种广泛的技能发展满足了MRC关于定量和跨学科技能的战略优先事项。

Programme overview: This MRC-funded doctoral training partnership (DTP) brings together cutting-edge molecular and analytical sciences with innovative computational approaches in data analysis to enable students to address hypothesis-led biomedical research questions. This is a 4-year programme whose first year involves a series of taught modules and two laboratory-based research projects that lead to an MSc in Interdisciplinary Biomedical Research. The first two terms consist of a selection of taught modules that allow students to gain a solid grounding in multidisciplinary science. Students also attend a series of masterclasses led by academic and industry experts in areas of molecular, cellular and tissue dynamics, microbiology and infection, applied biomedical technologies and artificial intelligence and data science. During the third and summer terms students conduct two eleven-week research projects in labs of their choice. Project:Clathrin-mediated endocytosis is one of the major pathways by which molecules are selectively imported into cells and plays a central role in a wide range of fundamental processes such as synaptic vesicle recycling, receptor recycling, signalling and development. This is achieved by formation of a proteinaceous clathrin coat, which is governed by interactions of clathrin with adaptor proteins, leading to vesicle budding and formation. Although our understanding is growing as to the roles of individual adaptor proteins during endocytosis, we still do not understand the manner in which multiple adaptor proteins interact with the clathrin cage or the molecular details of their interactions with one another.The two overarching aims are: 1. To investigate how key adaptor proteins affect the kinetics of assembly and disassembly using multiple biophysical techniques including dynamic light scattering, fluorescence spectroscopy and surface plasmon resonance.2. To carry out 3D structural analysis of clathrin cages bound to adaptors using high resolution cryo-EM and single particle analysis in order to visualise the mode of binding of key adaptors to assembled clathrin.During the studentship several skills from different disciplines will be developed, including how to conduct biophysical techniques such as surface plasmon resonance, dynamic light scattering, and fluorescence spectroscopy, and learn how to analyse the kinetic data obtained from these biophysical techniques. Structural and molecular biology techniques developed will include protein purification and expression, protein gel analysis (SDS-PAGE), GST pull downs, high resolution cryo-electron microscopy, and single particle analysis. This wide range of skills development fulfils the MRC strategic priorities for quantitative and interdisciplinary skills.