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

方案概览：这个由MRC资助的博士培训伙伴关系（DTP）将尖端的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，使学生能够解决以假设为主导的生物医学研究问题。这是一个为期4年的计划，其第一年涉及一系列教学模块和两个基于实验室的研究项目，导致跨学科生物医学研究硕士学位。前两个学期包括一系列教学模块，让学生在多学科科学中获得坚实的基础。学生还参加了一系列由学术和行业专家领导的大师班，这些专家在分子，细胞和组织动力学，微生物学和感染，应用生物医学技术以及人工智能和数据科学领域。在第三和夏季学期，学生在他们选择的实验室进行两个为期11周的研究项目。项目：网格蛋白介导的内吞作用是分子选择性输入细胞的主要途径之一，在突触囊泡再循环、受体再循环、信号传导和发育等广泛的基本过程中发挥核心作用。这是通过形成蛋白质网格蛋白外壳来实现的，这是由网格蛋白与衔接蛋白的相互作用控制的，导致囊泡出芽和形成。虽然我们的理解正在增长，以个别衔接蛋白在胞吞作用的作用，我们仍然不明白的方式，其中多个衔接蛋白与网格蛋白笼相互作用或它们相互作用的分子细节。利用动态光散射、荧光光谱和表面等离子体共振等生物物理技术研究关键衔接蛋白对组装和拆卸动力学的影响.使用高分辨率cryo-EM和单粒子分析对与接头结合的网格蛋白笼进行3D结构分析，以可视化关键接头与组装网格蛋白的结合模式。在学生期间，将培养不同学科的几项技能，包括如何进行生物物理技术，如表面等离子体共振，动态光散射和荧光光谱，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唐斯、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MRC的定量和跨学科技能的战略重点。

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.