HDeXperts

Hydrogen deuterium exchange mass spectrometry (HDX-MS) is used to measure the exchange rate of amide hydrogens with deuterated solvent. The exchange rate of amide hydrogens is exquisitely sensitive to changes in secondary structure and solvent accessibility, and this technique has been used extensively to characterize protein folding, as well as protein-protein, protein-lipid, and protein-small molecule binding sites.

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HDX-MS experiments can be carried out with a variety of stimuli, including protein binding partners, ligands, and membrane surfaces. Importantly, HDX-MS provides information on allosteric conformational changes. Our laboratory has made significant progress in the use of HDX-MS to study membrane localized signalling complexes that have been challenging to study by other structural approaches. We provide epitope mapping of antibody binding sites, and mapping of protein-protein interactions. We also provide screening of oncogenic specific drug binding interfaces.

 

In tandem with this work, we have found HDX-MS as a powerful tool in the optimization of crystallographic constructs, through the removal of intrinsically disordered segments. This is particularly useful in large multicomponent protein complexes, where disorder prediction software is of limited utility. Many proteins contain large intrinsically disordered regions, and it is highly unlikely for these regions to be observed in any high-resolution structural approaches. The structural mass spectrometry approach allows us to examine the role of intrinsically disordered regions in mediating interactions with other proteins, as well as in membrane binding, and protein function.