Cyclopropenones to assemble, analyze, and activate biomolecules

项目摘要 化学工具可以提供对生物分子结构和功能的独特见解。包括在此组中 是生物正交化学记者-生物兼容的官能团，可以针对不同类别的 并选择性地与各种探针连接。这些记者和反应使 生物分子将在他们的自然环境中进行分析。尽管他们过去取得了成功，并继续 潜在的，大多数化学记者已经很慢地过渡到生物研究和科学 整个社区。许多记者太大或不够稳定，无法正常使用手机，或者缺乏 化学多功能性。我们工作的长期目标是开发坚固耐用、易于使用的化学探针 用于跟踪和控制细胞和组织中的生物分子。此应用程序的目标是开发 一种试剂--环丙烯--作为一种通用的、用途广泛的化学报告。环丙烯酮港 独特的生物应用功能。它们很小，与生物正交性软亲核试剂反应。 (例如，磷化氢)，并且高度可调。除了标记生物分子外，还可以使用环丙烯酮 通过膦介导的交联和降解化学来控制生物分子的功能。因此，从一个 从理论上讲，单环丙烯单元可以组装、分析和激活感兴趣的生物分子。 这种试剂的多功能性和可及性可以为工具用户提供一站式服务，带来 化学探测器更快地进入非专业人员的手中。在强劲的初步数据指引下，我们的 工作将包括以下具体目标：1)调整环丙烯的广谱反应性 生物分子分析；(2)利用“潜在的”环丙烯反应性进行生物分子组装；以及(3) 建立生物分子功能的化学触发因素。在第一个目标下，我们会研究 环丙烯-膦连接，并鉴定具有合适稳定性和反应性特征的支架。在 第二个目标，我们将利用“潜在的”环丙烯反应来组装生物分子偶联物。 在第三个目标中，我们将开发可用于交联和激活的环丙酮触发剂 感兴趣的生物分子。我们的方法是高度创新的，因为它利用了一种独特的反应 获得多功能生物正交性的机制。拟议的研究具有重要意义，因为它 将提供多用途、易用的化学工具，适用于广泛的生物医学领域 研究。探测器还将使现有工具集无法进行的实验成为可能。另外，就像 对于其他化学技术，建议的试剂可能会在不同的领域激发新的发现。

Project Summary Chemical tools can provide unique insights into biomolecule structure and function. Included in this group are bioorthogonal chemical reporters—biocompatible functional groups that can target diverse classes of biomolecules and be selectively ligated with various probes. These reporters and reactions enable biomolecules to be analyzed in their native environments. Despite their past successes and continued potential, most chemical reporters have been slow to transition to biological studies and the scientific community at large. Many reporters are too large or insufficiently stable for routine cellular use, or lack chemical versatility. The long-term goal of our work is to develop robust, easy-to-access chemical probes for tracking and controlling biomolecules in cells and tissues. The objective of this application is to develop one reagent—cyclopropenone—as a general and versatile chemical reporter. Cyclopropenones harbor unique features for biological application. They are small, reactive with bioorthogonal soft nucleophiles (e.g., phosphines), and highly tunable. In addition to tagging biomolecules, cyclopropenones can be used to control biomolecule function via phosphine-mediated crosslinking and decaging chemistries. Thus, from a single cyclopropenone unit, one—in theory—can assemble, analyze, and activate biomolecules of interest. The versatility and accessibility of such reagents can provide a “one-stop-shop” for tool users, bringing chemical probes more rapidly into the hands of non-specialists. Guided by strong preliminary data, our work will encompass the following specific aims: 1) Tune cyclopropenone reactivity for broad-spectrum biomolecule analyses; (2) Exploit “latent” cyclopropenone reactivities for biomolecule assembly; and (3) Establish chemical triggers of biomolecule function. Under the first aim, we will examine the scope of the cyclopropenone-phosphine ligation, and identify scaffolds with suitable stability and reactivity profiles. In the second aim, we will capitalize on “latent” cyclopropenone reactivities to assemble biomolecule conjugates. In the third aim, we will develop cyclopropenone triggers that can be used to crosslink and activate biomolecules of interest. Our approach is highly innovative, as it capitalizes on a unique reaction mechanism to access multi-functional bioorthogonal properties. The proposed research is significant, as it will provide versatile, easy-to-use chemical tools that are applicable to a broad spectrum of biomedical research. The probes will also enable experiments not possible with existing toolsets. Additionally, like other chemical technologies, the proposed reagents will likely inspire new discoveries in diverse fields.