Characterisation and metrology
Cryo-electron microscopy structure determination
Cryo-electron microscopy has become a leading structural biology technique and Leeds offers world class facilities in this area. Not only does it house 2 Titan Krios microscopes but recent investments have provided the development of its tomography workflows with the purchase of a Hydra Bio Plasma FIB and recruitment of support staff. This allows us to now tackle a range of diverse scientific questions from purified proteins to in-situ studies of protein complexes. Furthermore, we have developed new technologies for time-resolved methodologies, sample preparation and tomography allowing us to be at the forefront of the field, not just purchasing the latest equipment but developing it to. Recent highlights have included the first in-situ study of amyloid fibrils within diseased tissue, trapping the myosin motor in action and developing molecular "glues". These studies have incorporated a large range of collaborations across academia and industry including projects with UCB Pharma, GlaxoSmithKline and Pfizer. An example high-resolution cryo-EM membrane protein structure is shown in Figure 1.
Figure 1. Structure of the quinol-dependent nitric oxide reductase AxqNOR by CryoEM. (a) CryoEM map of AxqNOR as determined by cryoEM to 2.2A (b) Dimeric AxqNOR in the plane of the bilayer represented by red lines with waters coloured as red spheres. https://doi.org/10.1038/s41467-023-39140-x (Flynn et al., 2023)
Key references:
Gilbert M.A.G., Fatima N., Jenkins J., et al.; CryoET of β-amyloid and tau within postmortem Alzheimer’s disease brain. Nature 631, 913–919 (2024).
Klebl D.P., McMillan S.N., Risi C., et al.; Swinging lever mechanism of myosin directly shown by time-resolved cryo-EM. Nature 642, 519–526 (2025).
Chandler F., Reddy P.A.N., Bhutda S., et al.; Molecular glues that inhibit deubiquitylase activity and inflammatory signaling. Nature Structural and Molecular Biology 32, 1812–1824 (2025).
Mass spectrometry
Leeds offers a state-of-the-art mass spectrometry centre which enables the characterisation of the structure and conformations of biomolecules and their complexes. The platform supports studies ranging from purified proteins to in-cell studies. The centre houses a Waters M-Class UPLC and Synapt G2S-i mass spectrometer for Hydrogen/Deuterium Exchange (HDX)-MS. By monitoring the exchange of backbone amide hydrogens with deuterium, HDX-MS provides site-resolved insights into protein folding, conformational changes, ligand binding, and protein–protein interactions under near-physiological conditions. These capabilities enable us to interrogate dynamic structural changes that are often inaccessible to static structural techniques alone. Recent highlights include the application of differential HDX-MS to probe conformational dynamics in the bacterial BAM complex and revealing how chaperones stabilise client proteins during folding.
Key references:
Csoma N., Machin J.M., Whitehouse J.M., et al.; Molecular insights into how the motions of the β-barrel and POTRA domains of BamA are coupled for efficient function. Nature Communications 16, 8832 (2025).
Wu K., Minshull T.C., Radford S.E., et al.; Trigger factor both holds and folds its client proteins. Nature Communications 13, 4126 (2022).