Applications of Click Chemistry in Glycobiology

点击化学在糖生物学中的应用

• 批准号: 7842018
• 负责人: Peng Wu
• 金额: $ 24.9万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-15 至 2012-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7842018
• 关键词: Agglutinins; Alanine; Aldehydes; Alkynes; Amino Acids; Anabolism; Azides; Binding; Binding Sites; Biochemical; Biological Response Modifier Therapy; Biomedical Research; Biopolymers; Biosensor; Canavalia; Carbohydrates; Carbon; Cell Communication; Cell surface; Cells; Chemicals; Chemistry; Collaborations; Consensus Sequence; Cysteine; Data; Dendrimers; Detection; Development; Devices; Disease; Endoplasmic Reticulum; Enzymes; Fc Immunoglobulins; Figs - dietary; Generations; Genetic; Genetic Transcription; Glycobiology; Goals; Grant; Green Fluorescent Proteins; Homologous Gene; Hydrazones; Hyperglycemia; Immunoglobulin G; In Situ; Incubated; Inflammation; Insulin Resistance; Intercept; Knowledge; Label; Lactose; Lectin; Length; Libraries; Life; Link; Malignant Neoplasms; Mammalian Cell; Mannose; Membrane Proteins; Mentors; Methods; Modification; Molecular; Oximes; Pathway interactions; Peptides; Pharmacologic Substance; Phase; Physical condensation; Physics; Plasmids; Platelet-Derived Growth Factor Receptor; Point Mutation; Polysaccharides; Post-Translational Protein Processing; Process; Production; Property; Protein Binding; Protein Glycosylation; Protein-Carbohydrate Interaction; Proteins; Reaction; Recombinant Proteins; Recombinants; Research; Series; Site; Specificity; Streptomyces coelicolor; Structure; Surface; System; Technology; Testing; Therapeutic; Therapeutic Intervention; Transcriptional Regulation; Transferase; Transistors; Translations; Transmembrane Domain; Work; adduct; base; biotin hydrazide; carbohydrate binding protein; carbohydrate biosynthesis; diabetes mellitus therapy; experience; expression vector; fluorophore; formylglycine; functional group; glycosylation; improved; inhibitor/antagonist; novel; prevent; professor; protein transport; research study; scaffold; single walled carbon nanotube; small molecule; therapeutic protein; tool; voltage

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.

细胞表面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 标志着癌症和炎症的发生。在细胞内，它们也可以调节转录、翻译 作为蛋白质贩运。然而，在描述聚糖功能的分子基础方面已经取得了进展。 相当慢。这部分是由于聚糖的生物合成与其他生物聚合物不同，既不 模板驱动也不在直接转录控制下。因此，常规的遗传和生化 阐明聚糖功能及其与疾病相关性的方法所获得的信息有限。 该项目的长期目标是实现点击化学——一套强大的、可靠的和选择性的 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，我将在两个新领域扩展我的研究 下一个授予期（ROO）的指示。目标 1 是发现/开发聚糖小分子抑制剂 使用酶模板原位点击化学来生物合成和加工酶。为了原理证明，我 选择0-β-N-乙酰葡糖氨基转移酶（OGT）作为第一个靶点。我打算开发片段库 将筛选 OGT 自组装抑制剂。鉴于 0-GlcNAc 过量的相关性 改变长期高血糖，进而引发胰岛素抵抗，这些化合物 开发的可能在糖尿病治疗中应用。目标 2 是拦截聚糖生物合成途径 带有生物正交官能团的合成非天然底物，例如叠氮化物和炔烃。在 同时，我还将开发基于点击化学的新选择性反应，用于后续检测 活细胞。