Applications of Click Chemistry in Glycobiology

点击化学在糖生物学中的应用

• 批准号: 7251049
• 负责人: Peng Wu
• 金额: $ 9万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 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 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7251049
• 关键词: Aldehydes; Alkynes; Anabolism; Antibodies; Azides; Binding; Biochemical Genetics; Biological Assay; Biomedical Research; Biomimetics; Biopolymers; Biosensor; Carbohydrates; Carbon; Carbon Nanotubes; Cell surface; Cells; Chemicals; Chemistry; Code; Compatible; Condition; Copper; Dendrimers; Detection; Development; Disease; Engineering; Enzymes; Eukaryotic Cell; Evaluation; Event; Genetic; Genetic Transcription; Glycobiology; Glycoconjugates; Glycoproteins; Goals; Grant; Habitats; Hyperglycemia; In Situ; Inflammation; Insulin Resistance; Intercept; Investigation; Knowledge; Lectin; Libraries; Life; Ligation; Link; Malignant Neoplasms; Mediating; Mentors; Metabolic; Methods; Modification; Molecular; Nucleic Acids; Oligosaccharides; Organism; Pathway interactions; Physiological; Polysaccharides; Process; Protein Glycosylation; Protein-Carbohydrate Interaction; Proteins; Reaction; Reagent; Relative (related person); Research; Research Personnel; Series; Site; Technology; Therapeutic; Therapeutic Intervention; Transcriptional Regulation; Transferase; Translations; Triazoles; Uridine Diphosphate; base; carbohydrate biosynthesis; catalyst; design; diabetes mellitus therapy; directed evolution; experience; functional group; glycosylation; glycosyltransferase; in vivo; inhibitor/antagonist; intracellular protein transport; molecular recognition; professor; programs; protein transport; research study; sensor; small molecule; sugar; tool

DESCRIPTION (provided by applicant): Cell surface glycans are major determinants of cell-cell and cell-matrix 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. The major objective of the first granting period (K99) is to initiate steps for applying click chemistry in studies of protein glycosylation in the lab of Professor Carolyn Bertozzi (the mentor). Aim 1 intends to use click chemistry-based bioorthogonal reactions to prepare homogeneous glycoproteins with therapeutic value. A genetically encoded aldehyde tag will be used for site-specific glycosylation. Aim 2 is to design and build specific lectin and antibody sensors using glycodendrimer-functionalized carbon nanotubes. With the experience and knowledge gained from the research in the first granting period, I will expand the applications of click chemistry in two new directions in the next granting period (R00). Aim 3 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 O-beta-N-acetylglucosaminyl-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 O-GlcNAc modification with insulin resistance-triggered hyperglycemia, the compounds developed may have applications in diabetes therapy. Aim 4 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. Glycans are known to participate in many normal and disease processes. This series of experiments will advance our understanding of glycan biosynthesis and carbohydrate-protein interactions related to these disease states. These studies may also offer new avenues for therapeutic intervention. I anticipate that the new chemical tools developed in these studies will have broad applications in biomedical research.

性状（由申请方提供）：细胞表面聚糖是细胞-细胞和细胞-基质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，它们可以调节转录，翻译以及蛋白质运输。然而，对聚糖功能的分子基础的描述进展相当缓慢。这部分是由于聚糖的生物合成与其他生物聚合物不同，既不是模板驱动的，也不是直接转录控制的。因此，用于阐明聚糖功能及其与疾病的相关性的常规遗传和生物化学方法产生的信息有限。该项目的长期目标是实现点击化学-一组强大，可靠和选择性的反应-作为糖生物学基础研究的通用工具。 第一个授权期（K99）的主要目标是启动在Carolyn Bertozzi教授（导师）实验室中将点击化学应用于蛋白质糖基化研究的步骤。目标1打算利用基于点击化学的生物正交反应来制备具有治疗价值的同质糖蛋白。基因编码的醛标签将用于位点特异性糖基化。目的二是利用糖树枝状聚合物功能化的碳纳米管设计并构建特异性凝集素和抗体传感器。 在第一个资助期的研究中获得的经验和知识，我将在下一个资助期（R 00）在两个新的方向上扩展点击化学的应用。目的3是利用酶模板原位点击化学发现/开发聚糖生物合成和加工酶的小分子抑制剂。为了证明原理，我选择O-β-N-乙酰葡糖胺基转移酶（OGT）作为第一个靶标。我计划开发片段库，筛选自组装的OGT抑制剂。鉴于过量的O-GlcNAc修饰与胰岛素抵抗触发的高血糖症的相关性，所开发的化合物可能在糖尿病治疗中具有应用。目的4是用合成的带有生物正交官能团（如叠氮化物和炔）的非天然底物阻断聚糖生物合成途径。同时，我还将开发基于点击化学的新的选择性反应，用于随后在活细胞中的检测。 已知聚糖参与许多正常和疾病过程。这一系列的实验将推进我们对与这些疾病状态相关的聚糖生物合成和碳水化合物-蛋白质相互作用的理解。这些研究也可能为治疗干预提供新的途径。我预计，在这些研究中开发的新化学工具将在生物医学研究中有广泛的应用。