Hydrogen deuterium exchange (HDX) is a powerful technique used to elucidate protein structures and their dynamics, thereby providing supporting information for X-ray crystallography and Cryo electron microscopy experiments. This technique is increasingly applied to diverse research questions and is becoming integral to advancing the study of structural dynamics in relation to basic biology and diseases. HDX utilises the exchange of labile hydrogen atoms on the solvent-exposed regions of proteins with deuterium. This exchange reaction is monitored over time mass spectrometry leveraging the distinct mass difference between deuterium and hydrogen. The rate of hydrogen deuterium exchange during HDX-MS is influenced by factors such as hydrogen bonding and solvent accessibility, offering valuable insights into protein dynamic change of the conformation. The applications of HDX-MS encompass protein unfolding, measurement of protein dynamic conformational changes, determination of the protein binding characteristics and interacting surfaces, epitope mapping, and identification of allosteric effects. HDX-MS presents notable advantages, particularly in the study of large protein complexes and membrane proteins that are challenges for other techniques.
At the Monash Proteomics & Metabolomics Platform, we have started to implement HDX-MS into our service repertoire, and we use Trajan Scientific's Parallel Extended HDX system attached to a Thermo Q Exactive HF Hybrid Quadrupole-Orbitrap mass spectrometer with Trajan Scientific's HDx-3 PAL™ platform. Our aim is to offer a full service to clients and collaborators ranging from experimental design to in-depth data analysis. To demonstrate the utility of HDX-MS, we have successfully applied this approach to cytochrome C as a model protein and the results align accurately with its known structure. Our team of experienced scientists is committed to delivering high-quality results and providing personalised support to our clients. Contact us to learn more about our HDX-MS services and how we can assist you in achieving your research objectives.