Unravelling the conformational dynamics of membrane proteins within a lipid environment
Dr Argyris Politis, Department of Chemistry, King's College London
Professor Sergi Garcia-Manyes, Department of Physics and Randall Division of Cell and Molecular Biophysics, King's College London.
Dr Kevin Giles, Corporate Scientific Fellow, Micromass (Waters Corporation)
This studentship will enable a new multidisciplinary strategy for elucidating the structural dynamics of membrane proteins in lipid context. Membrane proteins are embedded into a heterogeneous mixture of lipids that affects their structure, dynamics and function. Characterising protein-lipid interactions is a challenge and only a small portion of membrane protein structures is available in the relevant databases. Moreover, we know frustrating little about membrane protein dynamics, thus hindering a detailed understanding of their mechanism of action.
With this project, we will bring together a new combination of mass spectrometry (MS)-based approaches – hydrogen-deuterium exchange (HDX)-MS, and cyclic ion mobility (IM)-MS – with single molecule atomic force microscopy, AFM. HDX-MS will probe lipid-induced protein dynamics at peptide-level resolution, cyclic IM-MS will uncover co-populated states and unfolding pathways and AFM will elucidate conformational pathways and differences in (un)folding pathways.
The novelty lies in combining these techniques for probing lipid-modulated structural changes in membrane proteins; such changes are largely inaccessible by the existing approaches in isolation. We will establish our approach using exemplar membrane proteins for which high-resolution structures are available. We will examine how the surrounding lipid environment affects protein dynamics using nanodiscs. The nanodisc technology allows for reconstitution of membrane proteins into a native-like environment of tuneable lipid composition. Overall, the innovative integration of these tools will enable a new strategy for probing membrane protein dynamics and can readily be extended to any membrane protein of interest.
The student will be integrated into two dynamic and highly productive labs at King’s College London and an academic partner (Waters). They will be trained in expressing, purifying and reconstituting membrane proteins in nanodiscs as well as performing HDX-MS, IM-MS and AFM experiments. The student will spend time working at Waters Headquarters in Manchester where they will be part of vibrant industrial environment of a leading manufacturers of mass spectrometers worldwide.
eligibility and application
Applicants must hold, or be expected to achieve, a first or high upper second-class undergraduate honours degree or equivalent (for example BA, BSc, MSci) or a Masters degree in a relevant subject. This project is funded by a 4-year BBSRC studentship, applicants should ensure they have understood the funding eligibility criteria for these studentships. Unfortunately international students are not eligible for programme funding on this project.
The ideal candidate for this project would hold a Master’s degree in biochemistry, chemistry or related subjects. The candidate will have knowledge on molecular biology and protein chemistry. Expertise and interest in mass spectrometry and analytical science are desirable but not essential and not mandatory.
For more information regarding the project, please contact Dr Argyris Politis
Download the APPLICATION GUIDELINES here.