GLYCOPROTEIN GLYCOMICS: EXPRESSION IN MOUSE ES CELLS & PROGENY

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 Core 2A正在开发工具来分析细胞的糖蛋白糖化，包括胚胎小鼠干细胞及其分化产物。利用系列凝集素亲和层析分离胰酶糖肽，以根据所表达的糖的类型对表达进行分类。然后进一步用液相色谱分离糖肽，然后用质谱仪对各种物种进行编目和鉴定。在不同的阶段，N-葡聚糖酶处理将产生N-连接的葡聚糖。得到的多肽将通过LC/MS分析其天冬氨酸到天冬氨酸的转化来鉴定。所有结果将提交给生物信息学模块(核心项目4)，以便与其他数据整合。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Core 2A is developing tools to analyze the Glycoprotein Glycome of cells, including embryonic mouse stem cells and their differentiated products. Fractionation of tryptic glycopeptides using serial lectin affinity chromatography is being developed to classify expression by the types of glycans expressed. Separation of glycopeptides is then further accomplished by liquid chromatography, followed by mass spectrometry to catalog and identify various species. At various stages, N-linked glycans will be produced by N-glycanase treatment. The resulting peptides will be identified by their Asn to Asp conversion using LC/MS analysis. All results will be submitted to the Bioinformatics module (Core project 4) for integration with other data.