Research Techniques
Amide Hydrogen/Deuterium Exchange Mass Spectrometry (HDXMS)
Amide hydrogen/deuterium exchange (HDX), a powerful method to probe protein dynamics uses polypeptide backbone amide hydrogens as conformational probes. This exchange is dependent on structure (H-bonding), neighborhood effects and macromolecular interactions and is a readout of conformational flexibility in the millisecond-minute timescale. HDX-MS is especially useful as a comparative technique for probing conformational dynamics of proteins under biologically relevant perturbations including post-translational modifications, protein, lipid, nucleic acid and ligand interactions. Our group seeks to develop HDX-MS and ion mobility mass spectrometry methods and applications in protein dynamics.
Native mass spectrometry
(native-MS)
Native-MS is a useful method that measures the overall mass of intact proteins and protein complexes. By using electrospray ionization (ESI) on proteins in buffers such as ammonium acetate or ammonium bicarbonate, which readily ionize, the mass spectrometer can use the m/z values obtained to indicate protein folding, oligomeric states, and complex formation.
Cryo-Electron Microscopy (Cryo-EM)
Cryo-EM is a powerful method that enables us to get snapshots of proteins and virus particles in their native or perturbed environments. It begins by vitrifying protein samples on a cryo-EM grid and then analyzing using electron microscopy. When used in conjugation with HDX-MS and native-MS, protein and viral conformational shifts can be mapped on to static structures to obtain a fuller understanding of protein dynamics
