HDX-MS of membrane proteins
Our lab specializes in the use of Hydrogen-deuterium exchange mass spectrometry (HDX-MS) to probe conformation and dynamics of membrane proteins. We integrate a state-of-the-art mass spectrometry Bruker ultra high-resolution QTOF MS Impact II VIP (https://www.bruker.com/en/products-and-solutions/mass-spectrometry/qtof/impact.html) with Trajan LEAP Robotic system equipped with lipid filtration for studying membrane proteins in their most native environment. The combined instrumentation offers a robust system enabling automated analysis in a range of proteins and their complexes including membrane and soluble ones.
Membrane Protein Expression, Purification, and Reconstitution
HDX-MS provides a unique opportunity to investigate the dynamics of transporters, receptors and other membrane protein in various mimetic environments. Given the critical role of the lipid bilayer in stabilizing membrane proteins and influencing their functionality, studying these proteins in a native-like environment is essential. Although MS-friendly detergents have historically been preferred due to their ease of use and minimal interference with MS spectra, they are unable to accurately replicate the native lipid bilayer. As a result, there is a growing trend in HDX-MS research towards utilizing more native membrane mimetics, such as nanodiscs, liposomes, and membrane vesicles, to provide a more accurate representation of the protein’s natural environment.
In the Group, we are applying membrane reconstitution techniques with a focus on various mimetics to study transporter dynamics using HDX-MS. To best retain the native functionality and conformations of membrane proteins, our expression and purification processes involve bacterial and eukaryotic systems. Our lab and BSRC Fleming are well-equipped with essential instruments, including incubators for bacterial and insect cells, a cell sonicator, a cell disruptor, ultracentrifuges, and FPLC chromatography systems, to facilitate these critical procedures.
