Mass Spectral Sequencing of Carbohydrates

描述(由申请人提供)：上一个资金周期的努力已经启动了一种一致和独特的战略，用离子陷阱质谱仪(MSN)定义低聚糖的所有结构成分进行测序。为支持这一挖掘工作，开始了建立一个碎片资料库的工作，该资料库以符合HTP分析的数据确认、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、它们的立体化学和整体拓扑的详细信息。以相反的方式，这些路径是到前体离子的踪迹，已经为其编写了定义分子结构的代码。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选定的糖链拓扑结构。我们的研究正处于激动人心的时刻，但这一努力需要加强，需要更多和普遍适用的计算机工具来处理数据，以及更大一套合成标准和组织样本来扩大图书馆。建立一个寡聚体片段文库并编写更具体的代码是一项资源和人力的重大投资，但这是朝着确认的方向迈出的必要的下一步，也是我们迈向序列自动化的基础。。我们继续我们对应用的基本兴趣，不仅是作为原则的证明，而且是为了应对碳水化合物结构中要求苛刻的多营养参与。尤其具有挑战性的是已知的空间和时间表达模式，这些模式在细胞发育期间通过碳水化合物表位表现出来。为了解决这一研究领域，我们引入了一种新的、更合适的动物模型--水蚤。我们讨论了这种简单的微型甲壳动物适合于未来任务的许多原因，并提供了支持这一蛋白质组-血糖挑战的初步数据。水蚤以易驯化的、特征明确的生态环境和表现的表型，架起了基因与分子和细胞功能之间的桥梁。在合作中，作为水蚤基因组学联盟(致力于开发模型系统的国际研究人员网络)的成员，我们预计将支持一系列适合发育发现的基因KO和表型，可与我们现在对线虫所起的作用相媲美。

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.