My research has been mainly focused on the development of the novel mass spectrometry-based protein approaches for structural biology including currently a well-known and popular quantitative protein footprinting technology for examining large protein assembles. Large and dynamic multi-protein complexes control all critical cellular functions and understanding their physical organization is the key aim of structural biology. This novel protein footprinting method became a powerful tool to study protein structures and dynamics of protein assemblies and is widely utilized by many laboratories across the USA and the world.
Recently, I lead the development of a high-resolution MS-based hydroxyl radical protein footprinting approach to elucidate protein-protein/ligand binding interfaces. The level of single residues structural resolution which is provided by this method is essential for characterization of therapeutic antibody and protein-drug interactions, providing the drug-protein mechanisms assessment and guiding structure-based drug design.
However, a limitation of the current high-resolution footprinting methods is that labeling coverage using a single reagent with a single protease is still modest with ~5-20% of residues efficiently detected as labeled. My current efforts to overcome this limitation include development in collaboration with Drs. M. Gross and M. Chance of the novel footprinting chemistries suitable for use with the existing photolysis or proteolysis hydroxyl radical-based platforms. Specifically, by extending OH chemistry with simultaneous CF3 labeling we will enable a multiplex analysis to examine CF3 and OH related reaction events (which have different residue specificities) in the same experiment. These significant improvements in labeling chemistry will lead to substantially greater utility and impact for the technology in the important area of structural proteomics, with a focus on epitope/paratope mapping, characterization of large protein-protein complexes including amyloidogenic proteins and fibrils, membrane proteins including GPCRs and disordered proteins where other structural methods are currently insufficient to fully characterize them.