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

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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.