Applications of Click Chemistry in Glycobiology

细胞表面多聚糖是细胞间相互作用的主要决定因素。细胞表面糖基化的变化 标记癌症和炎症的发病。在细胞内，它们也可以调节转录和翻译 作为蛋白质交易。然而，在描述多糖功能的分子基础方面取得了进展 相当慢。这在一定程度上是因为，与其他生物聚合物不同，葡聚糖的生物合成既不是 模板驱动，也不受直接转录控制。因此，常规的遗传和生化 阐明多糖功能及其与疾病的相关性的方法产生的信息有限。 该项目长期目标是实现点击化学--一套强大、可靠和有选择性的 反应--作为糖生物学基础研究的通用工具。 凭借从K99阶段获得的经验和知识，我将在两个新的方面扩展我的研究 在下一个批准期(Roo)中的指示。目标1是发现/开发小分子糖链抑制剂 使用酶模板原位点击化学的生物合成和加工酶。为了证明原则，我 选择0-β-N-乙酰氨基葡萄糖转移酶(OGT)作为第一靶点。我计划开发片段库 这将被筛选出自组装的OGT抑制剂。给定过量的0-GlcNAc的相关性 在长期高血糖的情况下进行修饰，进而触发胰岛素抵抗，这些化合物 研发的药物可能会在糖尿病治疗中得到应用。目标2是通过以下途径截取多糖生物合成途径 合成的非天然底物，含有生物正交官能团，如叠氮化物和炔烃。在……里面 同时，我还将开发基于点击化学的新的选择性反应，以便随后在 活细胞。

Cell surface glycans are major determinants of cell-cell interactions. Changes In cell surface glycosylation mark the onset of cancer and inflammation. Inside the cell, they can regulate transcription, translation as well as protein trafficking. Progress toward delineating the molecular basis of glycan function, however, has been rather slow. This is partly due to the fact that the biosynthesis of glycans, unlike other biopolymers, is neither template-driven nor under direct transcriptional control. Therefore, conventional genetic and biochemical approaches for elucidating glycan function, and its relevance to disease, have yielded limited information. The long term goal of this project is to implement click chemistry-a set of powerful, reliable and selective reactions-as a general tool for fundamental studies of glycobiology. With the experience and knowledge gained from the K99 phase, I will expand my research in two new directions in the next granting period (ROO). Aim 1 is to discover/develop small molecule inhibitors of glycan biosynthetic and processing enzymes using enzyme-templated in situ click chemistry. For proof of principle, I chose 0-beta-N-acetylglucosamlnyl-transferase (OGT) as the first target. I plan to develop fragment libraries that will be screened for self-assembled inhibitors of OGT. Given the correlation of excessive 0-GlcNAc modification with prolonged hyperglycemia, which in turn triggers insulin resistance, the compounds developed may have applications in diabetes therapy. Aim 2 is to intercept glycan biosynthetic pathways with synthetic unnatural substrates bearing bioorthogonal functional groups, such as azides and alkynes. In parallel, I will also develop new selective reactions based on click chemistry for their subsequent detection in live cells.