The mass spectrometry facility runs routine HDX analyses to determine protein folding states, effects of ligand binding and protein interactions (e.g. epitope mapping). Further advanced instrumentation offers bespoke high-resolution native MS and ion mobility capabilities: Ultra-high mass range Q-Exactive Plus Orbitrap enabling native analysis of heterogeneous systems at high mass-to-charge ratios, e.g. membrane protein lipid complexes, and Tofwerk high-resolution ion mobility TOF (in late 2018) as well as several Waters Synapt HDMS systems.
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login to contact The Biomolecular Mass Spectrometry Facility provides access to the hydrogen/deuterium exchange (HDX)-MS capabilities. The instrumentation consists of LEAP sample handling robot, Waters UPLC with HDX Manager and Waters Synapt G2-Si. Data processing uses ProteinLynx Global Server and DynamX.
We perform peptide level analysis using online pepsin and aspergillopepsin proteolysis. Labelling durations can be chosen between 30s and 8h however in most cases 60 min should suffice as the maximum incubation time. Protein concentration is optimised during preliminary experiments. A minimum protein concentration of 10 µM and volume 150 uM being is required however higher stock concentrations are encouraged. Runs for each labelling condition takes approx. 16-18 hours depending on incubations chosen and the number of replicates. An additional day of access should be requested for instrument setup and preliminary experiments.