Incubation of protein samples in deuterated water with subsequent MS measurement can elucidate information pertaining to an analyte protein's higher-order structure. The rate of exchange of amide back-bone protons with deuterons in the bulk solvent is a function of solvent accessibilty and hydrogen bonding when termperature and pH are carefully controlled. These rates are affected by secondary, tertiary and quaternary structure and can be used to locate changes in protein conformation and dynamics as well as of binding interfaces between interacting partners.
Using Waters M-Class UPLC and Synapt G2S-i mass spectrometer purchased with funds from the BBSRC, we have developed robust protocols for perfoming comparative HDX-MS at the peptide level to probe a range of biologically relevant systems including protein-ligand interactions, nanobody & affimer binding and membrane proteins.
If you are interested in using HDX-MS in your research, or would like to find out more, whether you are part of the University of Leeds, other academic institutions or industry please get in touch.