Bioactive and bioorthogonal molecules for probing living systems

用于探测生命系统的生物活性和生物正交分子

• 批准号: RGPIN-2022-04234
• 负责人: Pezacki, JohnPaul
• 金额: $ 7.5万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=748199
• 关键词: Bioactive; bioorthogonal; molecules; probing; living

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.

我们的研究计划将继续推动现代化学生物学的边界。未来5年的计划将分为三个创新领域。首先，我们将继续开发新的生物正交化学，以允许修改和跟踪活的生物分子，包括来自人类病原体的蛋白质和RNA。我们的方法将集中在允许分子成像和识别的方法上。我们的小组是众所周知的开发环加成化学的生物正交标记应用的基础上的硝酮和其他兼容的功能团。最近，我们发现了新的化学方法，有望进一步应用和多重标记。这些涉及应变炔和烯烃与氧代吡啶离子的环加成，其具有相当水溶性和与生物流体相容的优点。我们还将继续在反电子需求狄尔斯-阿尔德化学中利用吡喃酮官能团取得进展。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组合，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掺入，所述非天然化学基团可用于询问引起特定病原性疾病的生物分子。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，促进有效的生物缀合反应在体内的应用。其次，我们将开发基于活性的蛋白质分析工具，以研究细胞和体内应用中酶活性的变化。基于活性的蛋白质谱（ABPP）是一种用于表征全细胞蛋白质组或活细胞复杂环境中酶功能的化学方法。这涉及到使用活性位点定向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，从而制备基于活性的探针。最后，我们将通过将非天然氨基酸掺入不同的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来赋予非天然蛋白质催化活性。该发现资助计划将继续进行基于自然科学化学原理的新化学生物学工具的创建工作。此外，这笔赠款将使我的实验室能够继续发展代表下一代加拿大科学家的有才华的多学科研究人员。这些高素质的人才将有助于加拿大未来在科学、技术和知识驱动的加拿大经济方面的竞争力。