New Chemical Strategies for the Modification and Control of Endogenous Proteins in Living Cells

修饰和控制活细胞内源蛋白的新化学策略

• 批准号: RGPIN-2014-04982
• 负责人: Menard, Frederic
• 金额: $ 2.33万
• 依托单位: University of British Columbia
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=659545
• 关键词: New; Chemical; Strategies; Modification; Control

Nature of the Research*Research in the Menard lab lies at the interface of chemistry and neurobiology. We seek to answer fundamental questions about how neurons communicate with their neighbouring cells, the astrocytes. This proposal describes the first steps we must take to achieve this goal. A long-term goal of our Chemical Biology research program is to create a chemical toolbox to study proteins that will complement the current genetic methods used by biologists. Specifically, we will: (1) design chemical tools to label proteins in live cells, and (2) apply these probes to understand the molecular mechanisms of synapse elimination in the aging brain. Our multidisciplinary research program integrates techniques of synthetic chemistry, molecular and cell biology, cell imaging, and electrophysiology. A central theme to this proposal is to modify natural products that have been evolved by Nature and use them as molecular probes to target specific proteins.**Significance*To better understand the mechanisms of neurodegeneration, scientists need new sets of tools to investigate protein-protein interactions at the molecular level. Having access to chemical means to monitor proteins without genetic modification would greatly accelerate the study of important cellular processes in health and disease. Current genetic methods to study protein dynamics suffer from significant drawbacks, e.g., protein expression or activity is often perturbed, and new vectors must be re-engineered for each organism. Chemists are uniquely positioned to design small molecules that can be applied at the time of experiment. This ensures we observe normal protein activity; moreover, in neurobiology, the molecules can easily be transferred from one organism to another due to the conserved nature of brain receptors across species. The successful implementation of our proposed methods will help unravel fundamental scientific questions at the frontiers of chemistry and biology. Such molecules will also be attractive tools for live cell assays in the pharmaceutical industry.**Expected Outcomes*This research proposal centers on the synthesis of small molecules that will offer novel means to visualize, label, and influence specific proteins in their native environment. Specifically, we expect this research will afford the chemical biology community with new reagents to achieve the traceless labeling of lysines residues in proteins. The broad applicability of the strategy of modifying natural products will demonstrate that new tools can be created from any ligand to study its associated protein in live cells. While we focus herein on cellular communication between neurons and astrocytes, the set of molecular tools we will use to study the calcium cascade will be applicable to any calcium-active tissue, such as heart cells and muscles. Finally, the systematic approach we will take to learn about chemical reactivity under physiological conditions should also lead to fundamental knowledge to help medicinal chemists design new drugs with covalent linkers (an emerging concept that can increase drug effiency several-fold).**Benefits to Canadians*Students and researchers in our lab will gain a unique training in interdisciplinary research including chemistry, molecular biology and neurobiology. While the field of chemical biology is growing rapidly, the expertise we bring is unique in Canada. In view of the pressing needs to treat neurodegenerative diseases, the students trained in our lab will be highly sought-after scientists at the national and international level. It is anticipated that the knowledge gained through these studies will provide valuable insight into how to develop pharmaceutical agents to treat or prevent neurodegenerative diseases.

研究性质 * 梅纳尔实验室的研究是化学和神经生物学的结合。我们试图回答有关神经元如何与相邻细胞（星形胶质细胞）通信的基本问题。这项建议描述了我们为实现这一目标必须采取的第一步。我们化学生物学研究计划的长期目标是创建一个化学工具箱来研究蛋白质，以补充生物学家目前使用的遗传方法。具体而言，我们将：（1）设计化学工具来标记活细胞中的蛋白质，（2）应用这些探针来了解衰老大脑中突触消除的分子机制。我们的多学科研究计划整合了合成化学，分子和细胞生物学，细胞成像和电生理学技术。该提案的一个中心主题是修改自然界进化的天然产物，并将其用作靶向特定蛋白质的分子探针。意义 * 为了更好地了解神经退行性变的机制，科学家需要一套新的工具来研究分子水平上的蛋白质-蛋白质相互作用。利用化学手段监测蛋白质而不进行基因改造将大大加快对健康和疾病中重要细胞过程的研究。目前研究蛋白质动力学的遗传方法存在明显的缺点，例如，蛋白质的表达或活性经常受到干扰，必须为每种生物体重新设计新的载体。化学家在设计可以在实验时应用的小分子方面具有独特的优势。这确保我们观察到正常的蛋白质活性;此外，在神经生物学中，由于跨物种的脑受体的保守性，分子可以很容易地从一个生物体转移到另一个生物体。我们提出的方法的成功实施将有助于解决化学和生物学前沿的基本科学问题。此类分子也将成为制药行业活细胞分析的有吸引力的工具。**预期成果 * 本研究计划的重点是小分子的合成，这将提供新的手段来可视化，标记和影响特定的蛋白质在其天然环境中。具体而言，我们希望这项研究将提供化学生物学社区与新的试剂，以实现蛋白质中赖氨酸残基的无痕标记。修饰天然产物的策略的广泛适用性将证明可以从任何配体创建新工具来研究活细胞中的相关蛋白质。虽然我们在本文中关注神经元和星形胶质细胞之间的细胞通信，但我们将用于研究钙级联的一套分子工具将适用于任何钙活性组织，如心脏细胞和肌肉。最后，我们将采取系统的方法来了解生理条件下的化学反应性，这也应该导致基础知识，以帮助药物化学家设计新的药物与共价连接（一个新兴的概念，可以提高药物效率几倍）。* 我们实验室的学生和研究人员将获得跨学科研究的独特培训，包括化学，分子生物学和神经生物学。虽然化学生物学领域正在迅速发展，但我们带来的专业知识在加拿大是独一无二的。鉴于治疗神经退行性疾病的迫切需求，在我们实验室接受培训的学生将成为国内和国际上备受追捧的科学家。预计通过这些研究获得的知识将为如何开发治疗或预防神经退行性疾病的药物提供有价值的见解。