Leveraging releasable aryl diazonium ions to probe biochemical systems

利用可释放的芳基重氮离子探测生化系统

• 批准号: 2320160
• 负责人: Hamish Christie
• 金额: $ 45.08万
• 依托单位: University of Arizona
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2320160&HistoricalAwards=false
• 关键词: Leveraging; releasable; aryl; diazonium; ions

With the support of the Chemistry of Life Processes (CLP) program in the Division of Chemistry, John Jewett from the University of Arizona is studying releasable reactive species to probe biological systems. Everything within biology can be analyzed based on its chemical composition. Reactive chemical probes form chemical bonds with molecules, both small and macromolecular-such as proteins or DNA, and these reactions are important because they can report back on the state of the system (healthy versus diseased, growing versus quiescent). Aryl diazonium ions (ADI) are a unique class of highly reactive molecules that the Jewett lab has learned to harness by rendering them deliverable using masking functional groups. These ADI could be leveraged to report on biological systems using fluorescence as an output to detect the chemical environment or enzymatic activity. The Jewett research team will also investigate the multifaceted fate of ADI once they enter a complex biological environment. In addition to their utility as biochemical tools, the study and synthesis of these compounds will serve to train undergraduate and graduate students. Moreover, this class of compounds generally exhibits fascinating color changes, fluorescence, solubility changes, and other properties that make them well suited to integrate with outreach goals. Dr. Jewett will develop an outreach program for a local school district with the goal of empowering elementary school students with safe experiments that they can carry out at home. Aryl diazonium ions (ADI) are powerful hypothesis-generating biochemical tools. A better understanding of ADIs is required to optimize their utility. One piece of this proposal seeks to develop a class of fluorogenic molecules with multiple mechanisms and inputs for activation. Multiple inputs could imbue these molecules with power to provide advanced spatiotemporal probing of the environment or of enzymatic status within a biochemical system. Additionally, the proposed research seeks to understand the nature of the reactivity and selectivity of ADI in living systems. While ADI have been used for decades for bioconjugation of proteins, or to degrade DNA, a holistic view of these disparate reactivities has not been undertaken. Following the synthesis of a small library of ADI, competition studies between disparate classes of biomolecules will be analyzed by NMR, mass spectrometry and gel electrophoresis. These studies could culminate in intracellular delivery of certain ADI using a triazabutadiene strategy. The findings of these studies may potentially impact the future of how this class of reactive probes is used for biological systems, from modifying purified proteins to probing whole organisms.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在化学系生命过程化学（CLP）项目的支持下，亚利桑那大学的John朱厄特正在研究可释放的活性物质，以探测生物系统。生物学中的一切都可以根据其化学成分进行分析。反应性化学探针与小分子和大分子（如蛋白质或DNA）形成化学键，这些反应很重要，因为它们可以报告系统的状态（健康与疾病，生长与静止）。芳基重氮离子（ADI）是一类独特的高反应性分子，朱厄特实验室已经学会通过使用掩蔽官能团使其可传递来利用这些分子。这些ADI可用于报告使用荧光作为输出的生物系统，以检测化学环境或酶活性。朱厄特研究小组还将调查ADI一旦进入复杂的生物环境后的多方面命运。除了作为生物化学工具的实用性外，这些化合物的研究和合成将用于培养本科生和研究生。此外，这类化合物通常表现出迷人的颜色变化，荧光，溶解度变化和其他特性，使它们非常适合与外展目标相结合。朱厄特博士将为当地一个学区制定一个推广计划，目标是让小学生能够在家里进行安全的实验。芳基重氮离子（ADI）是产生假说的有力生化工具。需要更好地了解ADI，以优化其效用。该提案的一部分旨在开发一类具有多种激活机制和输入的荧光分子。多个输入可以使这些分子具有能量，以提供对环境或生化系统内的酶状态的高级时空探测。此外，拟议的研究旨在了解ADI在生命系统中的反应性和选择性的性质。虽然ADI已被用于蛋白质的生物缀合或降解DNA数十年，但尚未对这些不同的反应性进行整体观察。在合成ADI的小型库之后，将通过NMR、质谱和凝胶电泳分析不同类别的生物分子之间的竞争研究。这些研究可能最终导致使用三氮杂环丁烷策略的某些ADI的细胞内递送。这些研究的结果可能会影响这类反应探针在生物系统中的应用前景，从修饰纯化蛋白质到探测整个生物体。该奖项反映了NSF的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。