text: It is not possible to determine the structure of a protein with H/D exchange other than neutron crystallography nor is it possible to define secondary structural elements.  The reasons for this are related to the way in which protein structure slows exchange.  Exchange rates are a function of two parameters: solvent accessibility and hydrogen bonding.  Thus an amide which is part of an intramolecular hydrogen bond will exchange slowly if at all, while an amide on the surface of protein hydrogen bonded to water will exchange rapidly. Amides buried from the solvent but not hydrogen bonded may also have very slow exchange rates.  Because both solvent accessibility and hydrogen bonding contribute to the rate of exchange, it becomes difficult to attribute a given exchange rate to a structural element without crystallography or NMR structural data.
  H–D exchange has been used to characterize the folding pathway of proteins, by refolding the protein under exchange conditions. In a forward exchange experiment (H to D), a pulse of deuterium is added after various amounts of refolding time.  The parts of the structure that form rapidly will be protected and thus not exchanged, whereas areas that fold late in the pathway will be exposed to the exchange for longer periods of time. Thus H/D exchange can be used to determine the sequence of various folding events. Factors determining the time resolution of this approach are the efficiency of mixing and how quickly the quench can be performed after the labeling.
  H–D exchange has been used to characterize protein structures and protein–protein interactions. The exchange reaction needs to be carried out with the isolated proteins and with the complex. The exchanging regions are then compared. If a region is buried by the binding, the amides in this region may be protected in the complex and exchange slowly. However, one must bear in mind that H–D exchange cannot be used to locate binding interfaces for all protein-protein interactions.  Some protein-protein interactions are driven by electrostatic forces of side chains and are unlikely to change the exchange rate of backbone amide hydrogens, particularly if the amide hydrogens are located in stable structural elements such as alpha helices.
  Lastly, H–D exchange can be used to monitor conformational changes in proteins as they relate to protein function.  If conformation is altered as result of post-translational modification, enzyme activation, drug binding or other functional events, there will likely be a change to H/D exchange that can be detected.