Bioactive and bioorthogonal molecules for probing living systems

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Our research program will continue to push the boundaries of modern chemical biology. The program for the next 5 years will be divided into three areas of  innovation. Firstly, we will continue to develop new bioorthogonal chemistry to allow the modification and tracking of living biomolecules including protein and RNA from human pathogens. Our approaches will be focused on methods that allow molecular imaging and identification. Our group is well known for developing cycloaddition chemistry for bioorthogonal labeling applications based on the nitrone and other compatible functional groups. Recently, we have discovered new chemistry that holds promise for further applications and multiplex labeling. These involve cycloadditions of strained alkynes and alkenes with oxopyridinium ions that have the advantage of being quite water soluble and compatible with biological fluids. We will also continue to make advancements with pyrone functional groups in reverse-electron demand Diels-Alder chemistry. Biological labeling will involve a combination of metabolic labeling and site specific modification using genetic code expansion, both of which are well established in our laboratory. The latter method involves the synthesis and incorporation of non-natural chemical groups that can be used to interrogate specific pathogenic disease causing biomolecules. Bioorthogonal reactions will be further developed to optimize reactivity in biologically relevant environments that promote efficient bioconjugation reactions for in vivo applications. Secondly, we will develop activity-based protein profiling tools to investigate changes in enzyme activity in cellular and in vivo applications. Activity-based protein profiling (ABPP) is a chemical approach for the characterization of enzymatic function within the complex environment of whole cell proteomes or living cells. This involves the use of active site-directed chemical probes that report on enzyme activity. We will design, synthesize and characterize new activity-based probes, with a special focus on polymerases and kinases, and related enzymes. We will also develop new reagents to quickly and conveniently modify natural products to make activity-based probes from. Lastly, we will develop new biocatalysts through the incorporation of unnatural amino acids into different proteins of interest. We will use unnatural metal-binding unnatural amino acids to impart catalytic activity to proteins that are unnatural. This discovery grant program will continue ongoing work in the creation of new chemical biology tools based on chemical principles in natural sciences. Furthermore, this grant will allow my laboratory to continue its development of talented and multidisciplinary researchers that represent the next generation of Canadian scientists. These highly qualified personnel will contribute to Canada's future competitiveness in science, technology, and our knowledge--driven Canadian economy.