Chemical Tools for the Investigation and Manipulation of Protein Glycosylation

摘要：“研究和操纵蛋白质糖基化的化学工具” 这项提议的广泛目标是开发化学工具，使 识别fi阳离子和抑制特定fic细胞的糖基化，糖基化介导的定位和细胞- 类型fic相互作用，以及碳水化合物生物合成途径的监测和操作。这个 在蛋白质中添加碳水化合物，或糖基化，是翻译后最常见的形式之一 Modifi阳离子，与各种过程有关，包括蛋白质稳定性、大分子 相互作用和细胞信号。不幸的是，目前可用来审问这些函数的工具 不足之处，这将糖基化的研究限制在专家实验室。我们计划通过以下方式解决这一未得到满足的需求 碳水化合物化学、光化学和化学生物学三种fic目标。在目标1中，我们将 建立在我们糖基化探针和抑制剂开发的基础上，重点是使用化学遗传 以细胞特有的fic方式创建工具来识别和干扰糖基化的方法。在目标2中，我们将 利用化学酶修饰fi阳离子糖基化的优点来安装特殊的fic光交联剂 到活细胞上，重点是识别由多糖介导的生物相互作用，以及 映射细胞间的相互作用。最后，在目标3中，我们将创建新的基于活动的探测器来测量和 抑制负责单糖生物合成的关键酶。在这些结束时 独立的目标，我们将产生新的强大的工具，将立即影响类型 科学家们可以就糖基化对人类健康和疾病的影响提出一系列问题。

ABSTRACT - “Chemical Tools for the Investigation and Manipulation of Protein Glycosylation” The broad goal of this proposal is the development of chemical tools that will enable the facile and robust identification and inhibition of glycosylation in specific cells, the mapping of glycosylation-mediated and cell- type specific interactions, and monitoring and manipulation of carbohydrate biosynthetic pathways. The addition of carbohydrates to proteins, or glycosylation, is one of the most common forms of posttranslational modifications and is associated with various processes, including protein stability, macromolecular interactions, and cellular signaling. Unfortunately, the currently available tools for interrogating these functions fall short, which limits the study of glycosylation to expert labs. We plan to tackle this unmet need using carbohydrate chemistry, photo-chemistry, and chemical biology in three specific aims. In Aim 1, we will build on our development of glycosylation probes and inhibitors, with a focus on using chemical genetic approaches to create tools to identify and perturb glycosylation in a cell-specific fashion. In Aim 2, we will leverage the advantages of chemoenzymatic modification of glycosylation to install specific photocrosslinkers onto living cells, with a focus on identifying biological interactions that are mediated by glycans, as well as mapping cell interactions. Finally, in Aim 3, we will create novel activity-based probes for measuring and inhibiting critical enzymes responsible for monosaccharide biosynthesis. At the conclusion of these independent aims, we will have generated new powerful tools that will have an immediate impact on the types of questions scientists can ask about glycosylation in human health and disease.