Function and regulation of epithelial glycosylation

摘要/摘要 这项研究计划的重点是揭示糖共轭化合物的生物学作用。这一努力的一部分是 致力于开发用于糖科学研究的化学生物学工具。在过去，我们发展了 光交联糖类似物，可结合到细胞糖偶联物中并用于共价 交联糖共轭在天然环境中与它们的结合伙伴结合。这些试剂可以用来鉴定 依赖于糖的结合作用，并表征这些结合在哪里和在什么条件下 相互作用就会发生。未来五年，我们将进一步扩大可以 通过制备额外的光交联糖，开发其掺入的新方法， 并评估它们与额外的糖共轭化合物的结合情况。使用这些光交联中的一种 糖，我们意外地观察到霍乱毒素可以与岩藻糖基化的糖结合在一起。 除了其典型的受体，神经节苷脂GM1。在未来五年，我们将确定 霍乱毒素识别的岩藻糖化糖偶联物的分子结构及其在霍乱毒素中的作用 宿主细胞中毒。这些研究得到了我们与Yrlid小组的长期合作的支持 (哥德堡大学)和他们在研究霍乱疾病机制方面的专门知识。我们对霍乱的研究 毒素受体使我们对排列在肠道和肠上的各种糖结合物感兴趣 呼吸道上皮细胞。一种CRISPR屏幕，旨在识别调节霍乱毒素与细胞结合的基因 表面鉴定了一些候选基因，这些基因可能通过不同的方式调节糖基化。 机械装置。在接下来的五年里，我们将描述糖基化的新调节剂，并确定如何 它们塑造糖链，调节糖链特征，如PolyLacNAc链长和 岩藻糖基化。这些研究的长期目标是确定不同个体之间的多糖特征是如何变化的， 它们与疾病状态的关联，以及它们对宿主-微生物相互作用的影响。

Summary/abstract This research program focuses on uncovering the biological roles of glycoconjugates. Part of this effort is devoted to the development of chemical biology tools for glycoscience research. In the past, we have developed photocrosslinking sugar analogs that can be incorporated into cellular glycoconjugates and used to covalently crosslink glycoconjugates to their binding partners in a native context. These reagents can be used to identify glycan-dependent binding interactions, and to characterize where and under what conditions that these interactions occur. Over the next five years, we will further expand the scope of experiments that can be performed by preparing additional photocrosslinking sugars, developing new methods for their incorporation, and evaluating their incorporation into additional glycoconjugates. Using one of these photocrosslinking sugars, we made the unexpected observation that cholera toxin can bind fucosylated glycoconjugates in addition to its canonical receptor, the ganglioside GM1. Over the next five years, we will determine the molecular structure of fucosylated glycoconjugates recognized by cholera toxin and characterize their role in host cell intoxication. These studies are supported by our long-term collaboration with the Yrlid group (University of Gothenburg) and their expertise in studying cholera disease mechanisms. Our studies of cholera toxin receptors led us to become interested in the diverse glycoconjugates that line the intestinal and respiratory epithelia. A CRISPR screen designed to identify genes that modulate cholera toxin binding to cell surfaces identified a number of candidate genes that may function in the regulation of glycosylation by diverse mechanisms. Over the next five years, we will characterize novel regulators of glycosylation and determine how they shape the glycome, modulating glycan features such as polyLacNAc chain length and the degree of fucosylation. The long-term goal of these studies is to determine how glycan features vary among individuals, their association with disease states, and their impact on host-microbe interactions.