MASS SPECTRAL SEQUENCING OF CARBOHYDRATES

描述（由申请人提供）：在过去的资助周期中，已经启动了一种一致和独特的策略，用于对寡糖进行测序，其中使用离子阱质谱仪（MSn）定义了所有结构组分。为了支持这一挖掘工作，开始了建立一个片段库的工作，该片段库以与HTP分析一致的数据确认、自下而上的方法吸收和检索光谱。碳水化合物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详细信息。以相反的方式，这些路径是前体离子的踪迹，已经为前体离子编写了代码以定义分子结构。这样的算法已经在不到一秒的时间内解决了新的和不寻常的结构上的选定聚糖拓扑结构。我们的研究正处于一个令人兴奋的时刻，但这一努力需要加强额外的和普遍适用的计算机工具，用于数据处理和一组更大的合成标准品和组织样本库扩展。建立寡聚体片段库和编写更具体的代码是资源和人力的重大投资，但它是确认的关键下一步，也是我们走向序列自动化的基础。.我们继续我们的应用程序的根本利益，不仅是作为一个原则的证明，但面对的碳水化合物结构的要求多营养参与。特别具有挑战性的是已知的空间和时间模式的表达，显示在细胞发育期间的碳水化合物表位。为了解决这一研究领域，我们介绍了一种新的和更合适的动物模型水蚤。我们讨论了许多原因，这种简单的微甲壳动物是适合未来的任务，并提供初步数据支持这一蛋白质组学的糖组学挑战。水蚤的桥梁基因和分子和细胞功能之间的跨度与听话和良好的特点生态和显示表型。通过合作，并作为水蚤基因组学联盟（致力于开发模型系统的国际研究者网络）的成员，我们期望支持一系列适合于发育发现的基因科斯和表型，与我们现在在C.优雅

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.