Bioactive and bioorthogonal molecules for probing living systems

我们的研究计划将继续推动现代化学生物学的发展。未来5年的项目将分为三个创新领域。首先，我们将继续发展新的生物正交化学，以允许修饰和跟踪来自人类病原体的活生物分子，包括蛋白质和RNA。我们的方法将集中在允许分子成像和识别的方法上。我们的团队以开发基于硝基和其他相容官能团的生物正交标记应用的环加成化学而闻名。最近，我们发现了新的化学物质，有望进一步应用和多重标签。这些方法包括用氧吡啶离子对张力炔和烯烃进行环加成，其优点是具有相当的水溶性和与生物流体相容。我们还将继续在吡酮官能团在反电子需求Diels-Alder化学方面取得进展。生物标记将涉及代谢标记和使用遗传密码扩展的位点特异性修饰的结合，这两者都在我们的实验室中得到了很好的建立。后一种方法涉及非天然化学基团的合成和结合，可用于询问特定的致病性疾病引起的生物分子。生物正交反应将进一步发展，以优化生物相关环境中的反应性，促进有效的生物偶联反应在体内的应用。其次，我们将开发基于活性的蛋白质分析工具来研究酶活性在细胞和体内应用中的变化。基于活性的蛋白质谱分析（ABPP）是一种在全细胞蛋白质组或活细胞的复杂环境中表征酶功能的化学方法。这包括使用活性位点导向的化学探针来报告酶的活性。我们将设计，合成和表征新的基于活性的探针，特别关注聚合酶和激酶，以及相关酶。我们还将开发新的试剂，以快速方便地修饰天然产物，使其成为基于活性的探针。最后，我们将通过将非天然氨基酸掺入不同感兴趣的蛋白质中来开发新的生物催化剂。我们将使用非天然金属结合非天然氨基酸赋予非天然蛋白质催化活性。这项发现资助计划将继续在基于自然科学化学原理的基础上创造新的化学生物学工具。此外，这笔拨款将使我的实验室能够继续培养有才华的多学科研究人员，这些研究人员代表了下一代加拿大科学家。这些高素质的人才将有助于加拿大未来在科学、技术和知识驱动型加拿大经济方面的竞争力。

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.