Mass spectrometry infrastructure for in-cell structural biology

The goal of this proposal is to establish a cutting-edge in-cell structural proteomics platform at the Astbury Centre that will enable the characterization of biomolecular structure and function in a cellular context.

This will inform on the architectures of proteins and protein complexes, including interactions with nucleic acids and small molecules, directly in their native environment.

We are requesting funding for the centrepiece of this platform, an Orbitrap Eclipse mass spectrometer with extended mass range and advanced fragmentation technologies.

This equipment will enable us to deploy the suite of structural proteomics methods we have developed for in vitro protein studies directly to the cell.

These approaches include in-cell covalent labelling to probe protein structure, such as by hydroxyl radical footprinting, and in-cell chemical crosslinking, including using bespoke photocrosslinkers, for interactome analyses.

Such methods can only be implemented utilising the most sensitive mass spectrometry instrumentation available, and require advanced fragmentation methods to achieve the highest possible structural resolution.

The proposed equipment will transform our ability to study molecular mechanisms of health and disease directly in the cell, and will result in translational opportunities, by identifying novel proteins, complexes and interactions that could be biomarkers or drug targets.