Structural Studies of Scaffords involved in Endocytosis and Signal Transduction

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The longterm goal of this project is to gain mechanistic understanding of how membrane remodeling and cytoskeletal rearrangements are orchestrated to culminate in the highly regulated process of endocytosis. To achieve this, I will solve the structure of a Bin/Amphiphysin/rvs protein (e.g. endophilin,amphiphysin or syndapin) protein with a well documented function in endocytosis. The protocols for expression and purification of these proteins are already established, leaving me time to focus on the structural aspects of the project. I will record electron cryomicroscopic images and use cutting-edge image processing strategies to reconstruct the structures of the membrane-bound proteins. While conceptually simple, the tasks at hand are not trivial and represent a boundary of what is technologically possible. Once structure determination of the first sample is underway, I will extend my efforts to purify interaction partners of the above-mentioned protein, like N-WASP, or dynamin (expression systems for these proteins have already been established locally), and pursue structure determination of these highly relevant complexes, preferably in a membrane-bound sate. The anticipated results will provide critical insights into the mechanisms of membrane remodeling, thus contributing to one of the most underrepresented fields in contemporary structural biology and biophysics.