Shotgun glycomics: linking glycan structure and function

鸟枪糖组学：连接聚糖结构和功能

• 批准号: 7814985
• 负责人: David Fletcher Smith
• 金额: $ 63.55万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-30 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7814985
• 关键词: Algorithms; Antibodies; Applications Grants; Award; Binding; Binding Proteins; Bioinformatics; Biological Assay; Blinded; Cell Line; Cells; Computing Methodologies; Data; Databases; Epitopes; Equipment; Funding; Glycoconjugates; Grant; HL-60 Cells; Knowledge; Laboratories; Lectin; Libraries; Link; Metadata; Methods; Molecular Biology; Nucleic Acids; Oligosaccharides; Organism; Parents; Plant Lectins; Polysaccharides; Printing; Process; Protein Binding; Proteins; Proteomics; Recovery; Research Personnel; Shotguns; Specificity; Structure; Surface; Testing; Tissues; Toxin; United States National Institutes of Health; computerized tools; nanoscale; novel strategies; public health relevance; response; tool

DESCRIPTION (provided by applicant): We are submitting this grant application in response to the NIH announcement of the Availability of Recovery Act Funds for Competitive Revision Applications as a supplement to the parent EUREKA Grant Award entitled, "Shotgun Glycomics: Linking Glycan Structure and Function." Shotgun Glycomics is a nanoscale approach that allows us to 1) fluorescently tag all free glycans released from cell or tissue glycoconjugates; 2) purify the glycans; and 3) create tagged glycan libraries (TGLs) that represent the glycome of the cell or tissue. Since glycans probably function as surfaces for effector molecules to bind, the tagged glycans are printed as glycan microarrays and interrogated for recognition by biologically relevant glycan-binding proteins (GBPs) or organisms. Sequencing oligosaccharides is extremely difficult and only done by highly specialized researchers with highly sophisticated equipment. Unlike nucleic acids and proteins, glycans cannot be amplified. Thus knowing sequences is of limited value. Shotgun Glycomics allows us to obtain an archival library of glycan structures on a microarray accessible to interrogation to gain knowledge about function so that we focus our sequencing and synthetic efforts on relevant glycans. As we developed TGLs and began interrogating them with GBPs, we recognized that massive amounts of structural information were available from the binding of GBPs with known specificity to unknown glycan structures. The exquisite specificity of plant lectins, for example, has been known and documented for decades. Thus, analyzing hundreds of isolated glycans on a microarray with a specific lectin could produce metadata for each glycan, and repeating the assay with additional lectins with different specificities rapidly increased the amount of metadata. We reasoned that if we included other parameters, we should be able to define glycan structures with simple microarray analyses. Thus, we are requesting support to develop a novel approach to glycan structure that we have termed "Metadata Analysis for Glycan Structure". This method only requires equipment common to molecular biology and proteomic labs, and we will develop computational tools and algorithms to convert metadata on glycan-protein binding to glycan structure. PUBLIC HEALTH RELEVANCE: Unlike nucleic acids and proteins, glycans cannot be amplified. Thus, knowing glycan sequences is of limited value. Shotgun Glycomics provides archival libraries of glycan structures for functional analysis. However, the same glycan array can be used to obtain structural data on all of the unknown glycan using "Metadata Analysis for Glycan Structure," a novel approach to glycan structure using microarrayed glycans, glycan specific proteins, and computational tools for metadata interpretation.

描述（由申请人提供）：我们提交此资助申请是为了响应NIH宣布的恢复法案资金的可用性，用于竞争性修订申请，作为对题为“猎枪糖组学：连接聚糖结构和功能”的尤里卡资助奖的补充。Shotgun Glycomics是一种纳米级方法，允许我们1）荧光标记从细胞或组织糖缀合物释放的所有游离聚糖; 2）纯化聚糖; 3）创建代表细胞或组织糖组的标记聚糖库（TGL）。由于聚糖可能作为效应分子结合的表面，标记的聚糖被打印为聚糖微阵列，并被生物学相关的聚糖结合蛋白（GBP）或生物体识别。低聚糖测序极其困难，只能由高度专业化的研究人员使用高度复杂的设备完成。与核酸和蛋白质不同，聚糖不能被扩增。因此，知道序列的价值有限。Shotgun Glycomics允许我们在可访问的微阵列上获得聚糖结构的档案库，以获得有关功能的知识，以便我们将测序和合成工作集中在相关聚糖上。当我们开发TGL并开始用GBP询问它们时，我们认识到大量的结构信息可从具有已知特异性的GBP与未知聚糖结构的结合中获得。例如，几十年来，人们已经知道并记录了植物凝集素的精致特异性。因此，用特异性凝集素分析微阵列上数百个分离的聚糖可以产生每个聚糖的元数据，并且用具有不同特异性的额外凝集素重复测定快速增加了元数据的量。我们推断，如果我们包括其他参数，我们应该能够用简单的微阵列分析来定义聚糖结构。因此，我们请求支持开发一种新的聚糖结构方法，我们称之为“聚糖结构元数据分析”。这种方法只需要分子生物学和蛋白质组学实验室常用的设备，我们将开发计算工具和算法，将聚糖-蛋白质结合的元数据转换为聚糖结构。 公共卫生相关性：与核酸和蛋白质不同，聚糖不能被扩增。因此，了解聚糖序列的价值有限。Shotgun Glycomics提供聚糖结构的档案库，用于功能分析。然而，相同的聚糖阵列可用于使用“聚糖结构的元数据分析”获得所有未知聚糖的结构数据，“元数据分析”是一种使用微阵列聚糖、聚糖特异性蛋白质和用于元数据解释的计算工具的聚糖结构的新方法。