HDX-MS as a Complementary Technique to Cryo-EM to Better Understand Protein Dynamics Upon Complex Formation or Small Molecule Binding Events

Philadelphia, April 26-28, 2023, NovAliX Conference “Biophysics in Drug Discovery”

Fabrice Ciesielski, Head of Protein Production and Structural Biology, shared some recent work of the NovAliX membrane protein team showing the recent structure determination of the GPCR TGR5 in complex with the G protein and a reference ligand P395. This is a great team achievement of the Structural Biology department.

Single particle cryo-electron microscopy (cryo-EM) and hydrogen/deuterium exchange mass spectrometry (HDX-MS) are both fast-growing structural biology techniques. Long seen as low-resolution techniques not able to compete with X-ray crystallography, both methods have benefited a lot over the last decade from methodological and IT improvements. HDX-MS is not only powerful for validating low-resolution cryo-EM structures but also for evidencing the dynamics of proteins. Due to their low consumption of protein, both techniques are particularly useful for a difficult to produce targets. Using a GPCR/G protein complex, G-protein coupled bile acid receptor 1 or TGR5, focus was given to the production and purification of the different proteins, complex formation and sample preparation for cryo-EM structure determination and HDX-MS analyses. We demonstrated how HDX-MS can reveal the dynamics of the proteins upon complex formation with the G proteins, and how the technique can be used to map the binding site of a small molecule or a nanobody.

This conference showed the very useful tool that HDX-MS can be to complement structural biology results.

Congratulations to the team for this great achievement!

Ieva, Marine, Clément, Alexandre, Thomas, Jérôme, Fabrice and Gilbert

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