Applications of Click Chemistry in Glycobiology

点击化学在糖生物学中的应用

基本信息

批准号：
7404452
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
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-04-15 至 2008-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7404452
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

项目摘要

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.

描述（由申请人提供）：细胞表面聚糖是细胞 - 细胞和细胞 - 矩阵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作。在细胞内部，它们可以调节转录，翻译和蛋白质运输。但是，朝着划定聚糖功能的分子基础的进展非常缓慢。这部分是由于以下事实：与其他生物聚合物不同，聚糖的生物合成既不是模板驱动的，也不是直接转录控制。因此，阐明聚糖功能及其与疾病相关的常规遗传和生化方法产生了有限的信息。该项目的长期目标是实施点击化学（一组强大，可靠和选择性的反应），作为糖生物学基础研究的一般工具。第一个授予期（K99）的主要目的是在Carolyn Bertozzi教授（导师）的实验室中启动在蛋白质糖基化研究中采用点击化学的步骤。 AIM 1打算使用基于化学的生物正交反应来制备具有治疗价值的均质糖蛋白。遗传编码的醛标签将用于特异性糖基化。 AIM 2是使用糖固定大分子官能化的碳纳米管设计和构建特定的凝集素和抗体传感器。凭借第一个授予期在研究中获得的经验和知识，我将在下一个授予期（R00）中将点击化学的应用在两个新的方向上扩展。 AIM 3是使用现场点击化学的酶发现/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，我选择了O-Beta-N-乙酰葡萄糖氨基转移酶（OGT）作为第一个目标。我计划开发碎片库，这些文库将用于OGT的自组装抑制剂。鉴于过度O-GLCNAC修饰与胰岛素抵抗触发的高血糖的相关性，开发的化合物可能在糖尿病治疗中有应用。目的4是用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）拦截聚糖生物合成途径。同时，我还将基于点击化学中的单击化学反应来开发新的选择性反应，以便它们在活细胞中的后续检测。已知聚糖参与许多正常和疾病过程。这一系列实验将提高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。我预计这些研究中开发的新化学工具将在生物医学研究中广泛应用。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Metabolic labeling of sialic acids in living animals with alkynyl sugars.

DOI：
10.1002/anie.200806319
发表时间：
2009
期刊：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
影响因子：
16.6
作者：
Chang, Pamela V.;Chen, Xing;Smyrniotis, Chris;Xenakis, Alexander;Hu, Tianshun;Bertozzi, Carolyn R.;Wu, Peng
通讯作者：
Wu, Peng

Boron nitride nanotubes are noncytotoxic and can be functionalized for interaction with proteins and cells.