Mass Spectral Sequencing of Carbohydrates

碳水化合物的质谱测序

• 批准号: 7268867
• 负责人: VERNON Nye REINHOLD
• 金额: $ 52.91万
• 依托单位: UNIVERSITY OF NEW HAMPSHIRE
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-01-01 至 2010-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7268867
• 关键词: Algorithms; Animal Model; Area; Automation; Biological Models; Caenorhabditis elegans; Carbohydrate Sequence; Carbohydrates; Cell physiology; Code; Collaborations; Commit; Computers; Daphnia; Data; Development; Diagnostic; Ecology; Epitopes; Funding; Genes; Genomics; Hexoses; International; Investments; Ions; Libraries; Mining; Molecular; Molecular Structure; N acetylhexosamine; Oligosaccharides; Pathway interactions; Pattern; Phenotype; Play; Polysaccharides; Proteomics; Publishing; Research; Research Personnel; Resources; Role; Standards of Weights and Measures; Structure; Tissue Sample; Work; Writing; carbohydrate structure; interest; man; mass spectrometer; member; monomer; stereochemistry; tool

DESCRIPTION (provided by applicant): The efforts of the past funding cycle have initiated a congruent and unique strategy for sequencing oligosaccharides with all components of structure defined using an ion trap mass spectrometer (MSn). In support of this mining effort, work was initiated to build a fragment library that assimilates, and retrieves spectra in a data-confirming, bottom-up approach consistent with HTP analysis. Carbohydrate MSn data and the pathways of disassembly provide details of monomer relationships, their stereochemistry, and overall topology. In a reverse manner these pathways are trails to the precursor ion for which code has been written to define molecular structure. Such algorithms have solved selected glycan topology, de novo, on new and unusual structures in less than one second. We are at an exciting point in our research, but this effort needs strengthening with additional and generally applicable computer tools for data handling and a larger set of synthetic standards and tissue samples for library expansion. Building a library of oligomer fragments and writing more specific code is a major investment of resources and man power, but it is an essential next step toward confirmation, and fundamental as we move to sequence automation. . We continue our fundamental interest in applications, not only as a proof of principle, but to confront the demanding pleotrophic involvement in carbohydrate structures. Particularly challenging are the known spatial and temporal patterns of expression that are displayed by carbohydrate epitopes during periods of cellular development. To tackle this research area we introduce a new and more appropriate animal model Daphnia. We discuss the many reasons this simple microcrustacean is appropriate for the tasks ahead and supply preliminary data supporting this proteomic-glycomic challenge. Daphnia bridges the span between genes and molecular and cellular functions with tractable and well-characterized ecologies and displayed phenotypes. In collaboration and as members of the Daphnia Genomics Consortium (an international network of investigators committed to developing the model system) we anticipate supporting an array of gene KOs and phenotypes appropriate for developmental discovery, comparable to the role we now play with C. elegans.

描述（由申请人提供）：过去的资助周期的努力已经启动了一种一致和独特的策略，用于测序低聚糖，其结构的所有组成部分使用离子阱质谱仪（MSn）定义。为了支持这种挖掘工作，开始建立一个片段库，以数据确认和自下而上的方法检索光谱，与HTP分析一致。碳水化合物的MSn数据和分解途径提供了单体关系、立体化学和整体拓扑结构的细节。反过来说，这些途径是通往前体离子的路径，而前体离子的代码是用来定义分子结构的。这种算法在不到一秒钟的时间内，就能在新的和不寻常的结构上重新解决选定的聚糖拓扑结构。我们的研究正处于一个激动人心的时刻，但这一努力需要加强，需要额外的、普遍适用的计算机工具来处理数据，还需要一套更大的合成标准和组织样本来扩展库。构建低聚物片段库和编写更具体的代码是对资源和人力的主要投资，但这是迈向确认的重要一步，也是我们走向序列自动化的基础。我们继续对应用的基本兴趣，不仅是作为原理的证明，而且是为了面对碳水化合物结构中要求的多营养参与。尤其具有挑战性的是在细胞发育期间碳水化合物表位所显示的已知的空间和时间表达模式。为了解决这一研究领域，我们引入了一种新的更合适的水蚤动物模型。我们讨论了这种简单的微甲壳类动物适合未来任务的许多原因，并提供了支持这种蛋白质组学-糖糖挑战的初步数据。水蚤在基因、分子和细胞功能之间架起了桥梁，具有易于处理和良好表征的生态和显示的表型。在合作中，作为水蚤基因组学联盟（一个致力于开发模型系统的国际研究人员网络）的成员，我们期望支持一系列适合发育发现的基因KOs和表型，可与我们现在在秀丽隐杆线虫中扮演的角色相媲美。