Grant: Discovery Grants Program - Individual

Our laboratory explores the mechanisms of protein folding and misfolding, as well as the relationship between protein dynamics, interactions, and function. For this purpose, we develop and apply novel electrospray ionization (ESI) mass spectrometry (MS) techniques that provide unique insights. We complement the resulting data with optical spectroscopy and cutting-edge molecular dynamics (MD) simulations. Past NSERC funding has allowed our group to move into a global leadership position. This DG will further extend the reach and scope of our activities by focusing on three interconnected themes.******1. Protein Dynamics and Function. Hydrogen/deuterium exchange (HDX)/MS is an important tool for probing protein dynamics in solution. Flexible segments undergo rapid HDX, while deuteration in folded regions is slow. We will develop a greatly improved understanding of HDX protection effects, providing the foundation for HDX/MS-based structural modeling efforts. HDX/MS will uncover the folding mechanisms, internal dynamics, and degradation phenomena of key proteins, including therapeutic antibodies. HDX/MS studies on protein-based molecular machines will elucidate aspects related to lubrication, friction, deformation, and power transmission. These results will be relevant for future applications in nanotechnology.******2. Analytical and Biomedical Aspects of Protein Oxidation. Innovative radical chemistry will overcome limitations of currently existing oxidative labeling methods, paving the way towards unprecedented insights into the structures of protein folding intermediates. We will also elucidate the role of protein oxidation for the peroxidase activation of cytochrome c, a topic that is related to the apoptotic elimination of cancer cells.******3. ESI Mechanisms and Gas Phase Proteins. “Native” ESI-MS is widely used for probing proteins and biological complexes. However, it remains unclear to what extent electrosprayed analytes retain their structures in the gas phase. We will scrutinize the hypothesis that extensive salt bridge networks impart kinetic stability to native-like gas phase proteins. In addition, our work will reveal whether proteins can fold in the gas phase, shedding light on the role of water during biomolecular self-assembly. Novel computational avenues will yield “molecular movies” of the ESI process, placing the field of native ESI-MS on solid theoretical/computational footing.******HQP Training. This proposal revolves around a carefully planned and executed HQP training strategy. Being part of our team provides HQP with an exceptional track record that includes high profile publications, major awards, and prestigious scholarships. Collaborations with other research groups and institutions greatly contribute to a stimulating interdisciplinary environment. This training background provides a springboard for prosperous careers in industry or academia.**