Enabling Technologies to Characterize Biomolecules and Fluorophores in Controlled Micro-environments using Mass Spectrometry

点击翻译按钮获取中文摘要

The Jockusch laboratory aims to create transformative tools and technologies that will have long-term impact on fields ranging from chemistry and physics to biology and medicine. This research program will advance understanding of how key molecules, such as proteins and DNA, are affected by their molecular environment. We primarily focus on building enabling technologies for mass spectrometry (MS), many of which exploit laser spectroscopy. Our work addresses critical challenges at heart of chemistry, biology and medicine. For example, proteins that are `misfolded' have costly impacts on health, due to their association with devastating diseases including Alzheimer's, Parkinson's and type II diabetes. Our contribution to combatting such protein-misfolding diseases is to develop and apply techniques that illuminate how different interactions contribute to protein folding, misfolding and aggregation. This will uncover interactions important for the development of efficacious drugs. In a related endeavor, we seek to improve the understanding of popular molecular sensors whose fluorescence is used to characterize key molecular states, including disease-correlated states. For example, we target understanding whether the best-known sensors for amyloid fibrils, which are late-stage misfolded protein aggregates, can report on the earlier-stage oligomers that are important therapeutic targets in Alzheimer's research. In order to dissect the intricate web of molecular interactions present in Nature, we begin by examining simplified systems whose complexity is then gradually built up in a carefully controlled fashion. We establish baseline behavior by characterizing key properties of bare, mass-selected ions. Then, pieces of the molecular environment (such as water molecules, metal ions, drugs or other binding partners) are added to form complexes and clusters, thus enabling discovery of how additional specific interactions modulate molecular properties of interest. Over the next five years, we aim to expand the suite of tools that can usefully be employed for molecular characterization. We primarily concentrate on developing new dimensions of readily-interpretable information from tandem MS analyses. For example, we made a flexible interface for laser spectroscopic experiments that enables the use of fluorescence and Förster resonance energy transfer (FRET) techniques to probe conformation, and other functional characteristics, of mass-selected ions. We also develop orthogonal probes of molecular characterization, such as gas-phase hydrogen-deuterium exchange. By combining information from these complementary techniques, a better picture will emerge of how molecular properties change with the local molecular environment. Overall, the techniques and tools developed in the Jockusch lab will be used to push scientific boundaries. This cutting-edge research program also provides an excellent training environment for the next generation of Canadian Scientists.