Grant: Discovery Grants Program - Individual

Most proteins in living organisms can perform their biological functions only after folding into a highly unique structure. Folding occurs spontaneously, driven by free energy gradients. Notably, the biologically active state remains quite dynamic, with structural fluctuations that are closely related to function. Misfolding can lead to Alzheimer's and many other diseases. Our group deciphers the mechanisms of protein folding and misfolding, as well as the interplay between structure, dynamics, and function. We have demonstrated international leadership in the development and application of novel protein mass spectrometry (MS) techniques. The current DG extends the reach and scope of these activities in a very significant way. (a) We will devise a method for elucidating the structures of the earliest folding intermediates. The combination of diffusive mixing with laser-induced oxidative labeling will improve the time resolution by three orders of magnitude compared to existing technologies. (b) We will employ oxidative labeling for mechanistic studies on a wide range of biologically important folding and self-assembly processes. (c) Hydrogen/deuterium exchange (HDX)/MS will be applied for gaining detailed insights into protein-ligand interactions. (d) Folding and dynamics of membrane proteins represent frontier areas that we will tackle using HDX/MS and oxidative labeling. (e) Shifting from traditional single-protein investigations to a ""Systems Biology"" perspective, we will be the first to apply structural/functional MS methods to protein networks, such as photosynthetic complexes. (f) Investigations on proteins in a solvent-free environment can reveal ""intrinsic"" biomolecular properties. Using a unique combination of gas-phase HDX, electron capture dissociation, ion mobility spectrometry, and computational modeling we will elucidate protein structures in the vacuum. Key to the success of all these activities is a carefully planned and executed strategy for HQP training. By becoming an integral part of our research enterprise HQP can develop an outstanding publication track record, as well as acquire the skills needed for successful careers in industry and academia.