Acquisition of a Waters Hydrogen/Deuterium eXchange (HDX) Mass Spectrometry System

This proposal is for the acquisition of a Waters Hydrogen/Deuterium eXchange (HDX) mass spectrometry system. Until recently, HDX technology required laborious manual handling and analysis, but new technological innovations have now made this process automated and streamlined. Currently, in order to currently perform HDX experiments, UK faculty must either partner with one of the few academic collaborators throughout the US or pay a company. These experiments are quite costly and even academic collaborators require $25,000-$35,000/annually to perform these experiments. The requested system is uniquely suited and specifically configured and optimized to carry out HDX studies, consisting of an HDX management module, Acquity UPLC, and SYNAPT G2-Si high definition mass spectrometer with ion mobility and electron transfer dissociation (EDT) capabilities. Obtaining this equipment will make this new technology broadly available to investigators at the University of Kentucky and significantly advance the research mission of the University. The PI has published extensively using HDX experiments, as have a number of major users. The co-PI has the expertise and technical assistance to incorporate HDX into the facility. It is also important that a significant number of the major and minor users are NSF funded, which will be a major criterion during review, and they are spread throughout five Colleges. Additionally, multiple faculty at Primarily Undergraduate Institutions (PUI) have expressed interest in accessing the system, and will be included as collaborators, significantly enhancing the broader impacts of the proposal. HDX is a powerful cutting-edge technique that can yield insights into the structure, dynamics, physical association, and/or specific binding of a given protein or macromolecule based on assessing the status of whether a peptide is surface exposed. Proteins in physiological buffers will exchange solvent accessible protons with deuterium. A change in the rate or number of protons exchanged with deuterium suggests a change in protein conformation. HDX mass spectrometry utilizes the mass difference between a hydrogen and a deuterium atom to measure the rate, number, and location of exchangeable hydrogen atoms in a protein of interest. HDX is not a traditional proteomic technique, instead it provides critical insights into the conformational dynamics in proteins, protein-ligand and protein-protein complexes.