Developing high affinity ligands of glycan binding proteins

开发聚糖结合蛋白的高亲和力配体

• 批准号: 9166443
• 负责人: Corwin Michael Nycholat
• 金额: $ 39.57万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9166443
• 关键词: Address; Affinity; Alkynes; Animals; Azides; Binding; Binding Proteins; Biological Assay; Carbohydrates; Cell-Matrix Junction; Cells; Chemicals; Chemistry; Computer Simulation; Copper; Development; Epitopes; Family; Flow Cytometry; Galactose; Galactose Binding Lectin; Galectin 3; Glycolipids; Glycoproteins; Heart Diseases; Human; Immune response; In Vitro; Infection; Inflammation; Lead; Libraries; Ligands; Link; Malignant Neoplasms; Mediating; Methods; Microspheres; Mus; N-acetyllactosamine; Pathologic Processes; Physiological Processes; Polysaccharides; Printing; Reaction; Reagent; Recombinants; Regulation; Research Personnel; Role; Signal Transduction; Site; Specialist; Structure; Testing; Therapeutic; Tissues; analog; base; combinatorial; cycloaddition; extracellular; in vivo; inhibitor/antagonist; microbial; screening; tool

PROJECT SUMMARY/ABSTRACT Glycan-binding proteins (GBPs) decode the dense structural information contained in the glycome produced by all cells. Galectins are a family of GBPs widely distributed among animals, which are expressed in various tissues, and occur in both intracellular and extracellular compartments. The carbohydrate recognition domain (CRD) of galectins bind to β-galactose epitopes such as N-acetyllactosamine (LacNAc) units commonly found on glycans of N- and O-linked glycoproteins, and glycolipids. Galectins mediate many physiological processes including cell-cell and cell-matrix adhesion, cell signaling, and regulation of immune responses. However, galectins are also implicated in pathological processes such as cancer, inflammation, heart disease, and regulation of microbial infection. The objective of this project is to develop high affinity and selective ligands of galectins based on glycan analogs that can block their interactions with glycans to provide valuable tools for investigating the functions of this important class of glycan-binding protein or serve as potential therapeutics. In Aim 1 we will generate a large diverse library of glycan analogs using several high- throughput strategies, including a combinatorial approach involving an on-chip click-chemistry reaction, and an in silico guided screen of galectin co-crystal structures to identify favorable substituents from large commercial building block libraries. The glycan analog library will be incorporated into a glycan microarray which will be screened in Aim 2 against a complete panel of human and murine galectins to identify high affinity and selective ligands. The printed array is readily adaptable and will be made available to other researchers including non-specialists for glycan array analysis. The galectin inhibitors developed in this project will be truly off-the-shelf reagents that non-specialists can easily adapt for functional studies or therapeutic applications.

项目摘要/摘要 聚糖结合蛋白（GBP）解码了所有细胞产生的Glyce中包含的致密结构信息。甲状腺激素是在动物中广泛分布的Gbps家族，它们在各种组织中表达，并发生在细胞内和细胞外室中。半乳胶状素的碳氢识别结构域（CRD）与β-半乳糖表位结合，例如N-乙酰乳糖胺（LACNAC）单位（LACNAC）单位，通常在N-和O-连接链染料的Glycans上发现的许多物理过程介导了许多物理过程，包括许多细胞 - 细胞 - 细胞 - 细胞 - 细胞和细胞 - MATRIX粘合剂，和细胞信号，和监管的细胞 - 和细胞信号。然而，在癌症，感染，心脏病和微生物感染的调节等病理过程中也暗示了甲状腺素。该项目的目的是基于聚糖类似物来开发高亲和力和选择性配体的甘叶蛋白，这些配体可以阻止其与聚糖的相互作用，以提供有价值的工具来研究这种重要类型的聚糖结合蛋白或作为潜在疗法的功能。在AIM 1中，我们将使用多种高通量策略生成一个大型的聚糖类似物库，包括涉及芯片咔嗒声反应的组合方法，以及在塞拉克蛋白共晶体结构的硅基指导屏幕中，以识别来自大型商业建筑块的有利的子托管。聚糖模拟库将被合并到一个聚糖微阵列中，该库将在AIM 2中与完整的人和鼠乳肠蛋白组成，以识别高亲和力和选择性配体。印刷阵列很容易适应，并将提供给其他研究人员，包括非专家进行聚糖阵列分析。该项目中开发的半乳糖素抑制剂将是真正的现成试剂，非专家可以轻松适应功能研究或治疗应用。