GLYCOPROTEINS EXPRESSING GLYCANS THAT REACT WITH DBA AND ANTI-POLY SIALIC ACID

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 干细胞计划(道尔顿)已经确定了消失的ES细胞上的扁豆凝集素(DBA)阳性表位 当诱导分化为EPL和EBOD时。DBA凝集素印迹鉴定6-10个糖蛋白表达糖链 由DBA识别。表达这些多糖的糖蛋白通过切下凝胶切片、胰酶消化、 MALDI-TOF MS分析。此外，这些糖蛋白上的葡聚糖被释放，并被吡啶胺标记，以及 经层析分离和MALDI-TOF MS鉴定，初步结果表明DBA表位是N-连接的 由于多聚糖对N-多聚糖释放的敏感性。最终，我们将回答DBA糖蛋白是如何 分化后反应性消失。 糖转录组计划(MOREMEMEN)已发现聚唾液酸合成酶在分化(EBOD)中的表达 细胞。EBOD和NOT ES细胞的聚唾液酸特异性抗体染色证实了这一结果。我们将确定 糖蛋白表达这种糖链的糖蛋白及其功能

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The Stem Cell Program (Dalton) has identified Dolichos biflorus agglutinin (DBA) positive epitopes on ES cells that disappear when differentiation to EPL and EBods is induced. Lectin blotting with DBA identified 6-10 glycoproteins expressing glycans recognized by DBA. The glycoproteins expressing these glycans are being identified by excision of gel slices, trypsinization, and MALDI-TOF MS analysis. Moreover, the glycans on these glycoproteins are released, tagged with pyridylamine, and identified by chromatographic separation and MALDI-TOF MS. Initial results suggest that the DBA epitopes are on N-linked glycans because of their sensitivity to N-glycans release. Ultimately, we will answer the question of how the DBA glycan reactivity disappears after differentiation. The Glycotranscriptome Program (Moremen) has identified polysialic acid synthase expression in differentiated (EBod) cells. Staining of EBod and not ES cells with antibody specific to polysialic acid confirms this result. We will identify the glycoproteins that express this glycan and their function.