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

衔接蛋白在网格蛋白外壳形成和分解动力学中的作用

基本信息

批准号：
1935264
负责人：
金额：
--
依托单位：
University of Warwick
依托单位国家：
英国
项目类别：
Studentship
财政年份：
2017
资助国家：
英国
起止时间：
2017 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=studentship-1935264
关键词：
role adaptor proteins dynamics clathrin

项目摘要

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.

计划概述：这种由MRC资助的博士培训合作伙伴（DTP）通过数据分析中的创新计算方法汇集了尖端的分子和分析科学，以使学生能够解决假设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授的模块和两个基于实验室的研究项目，这些项目导致跨学科生物医学研究的硕士学位。前两个术语包括一系列教学的模块，使学生能够在多学科科学中获得扎实的基础。学生还参加了一系列由学术和行业专家领导的大师班，该领域在分子，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以及数据科学领域。在第三和夏季期间，学生在他们选择的实验室中进行了两个11周的研究项目。项目：网格蛋白介导的内吞作用是将分子选择性进口到细胞中的主要途径之一，并在广泛的基本过程中起着核心作用，例如突触囊泡回收，受体回收，信号传导和发育。这是通过形成蛋白质网状蛋白外套来实现的，该蛋白质层由网格蛋白与衔接蛋白的相互作用控制，从而导致囊泡发芽和形成。尽管我们对内吞作用期间个体适配蛋白的作用的理解正在增长，但我们仍然不了解多种畸形蛋白与网状蛋白笼或它们相互作用的分子细节相互作用的方式，它们彼此之间的相互作用的分子细节，两个总体目的是：1。要调查如何使用多种型号的动态蛋白来构成多种型号的动态，以使组合的动态群体（流行型的动态）群体群体群体（流动性的动态），以使组合蛋白质群体群体群体群体群体（流行型的动态）群体群体群体群体群体群体群体群体群体群体群体，光谱和表面等离子体共振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从这些生物物理技术获得的数据。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下降，高分辨率的低分子电子显微镜和单个颗粒分析。这种广泛的技能发展履行了MRC战略优先事项，以实现定量和跨学科技能。