Recent biomedical advances including anti-viral and anti-cancer immunotherapies, and structure-based drug design are driven by insights from structural biology. Despite notable structure-based successes, membrane proteins remain difficult targets and consequently a significant challenge. Membrane proteins are comparatively high-risk, expensive and require significant optimisation for success. The Membrane Protein Laboratory (MPL) at Diamond Light Source assists users from all over the world with the expression, purification and structural characterisation of membrane proteins. Membrane proteins are ubiquitous in all organisms from prokaryotes to eukaryotes and play key structural and functional roles. In prokaryotes, membrane proteins are attractive targets for new antimicrobial therapies, whilst in eukaryotes, such as humans, understanding the structure and function of membrane proteins is important to understanding the molecular basis of diseases.
Outside of a cell, membrane proteins must be maintained in a hydrophobic environment. As such to study them biochemically and structurally we remove them from membranes into detergents (soaps). To help us in this process we have developed a series of high-throughput screens that allow us to identify optimal buffer, detergent or detergent-free systems. These screens accelerate the transfer of proteins from initial biochemical studies to structural studies and filter out poorly behaved samples before we attempt to structurally characterise them by techniques such as crystallisation or cryo-electron microscopy.
By removing membrane proteins from their native environment we lose opportunities to investigate the effect of cellular lipids on the native state of a membrane protein. We can partially restore the native state by incorporating our membrane protein into a liposome (essentially balls of lipids, like a micelle). Liposomes more closely resemble the native membrane and allow use to define the lipid content, enabling the effect of different lipids on structure and function to be studied. Ultimately, we would like to study membrane protein structure and function at atomic resolution in the cell but how do you identify your favourite protein from the thousands of others? One possible approach is to use small antibody fragments called nanobodies. The use of nanobodies in structural biology has been transformative in trapping protein conformations and enabling atomic resolution structures of membrane proteins. The unique structure of a nanobody enables them to access cavities within proteins such as the active sites of enzymes or the clefts of a receptors. These properties make them ideal tools to probe protein structure and function or even as potential biomedicines. By labelling nanobodies with a fluorescent probe we hope to identify our protein(s) of interest firstly in proteoliposomes and subsequently on the surface of cells and organelles.
Finding the right nanobody is an interactive process and as such well suited to the high-throughput screening platform we already use for membrane proteins. We plan to adapt our screening platform to rapidly identify the best nanobody binders as well as automate the platform for all of our existing screens using automated magnetic particle separation technology. Purified nanobodies will be labelled and we will use these to detect membrane protein reconstituted into proteoliposomes and on the surface of cells.
Applications are now closed for this project.
Please apply via our online application portal. The vacancy that you are applying for is the "Summer Placements 2023" listing, you will then have the opportunity to select up to three projects to apply for. This project's reference is 23001SP.
Interviews will be scheduled for 6, 7, 8, and 9 February 2023.
If you are disabled and would like to be considered under the Disability Confident Scheme, please let us know via the online application process.
Please note that this role does not meet the required skill level for a Skilled Worker visa and therefore we would be unable to sponsor individuals due to the current UK Home Office immigration rules. To be appointed to the role, candidates will need to have the right to work in the UK without sponsorship from us.
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