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(Dalton)发现了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阳性表位消失。凝集素与DBA杂交鉴定出6-10个表达DBA识别的糖蛋白的糖蛋白。表达这些糖蛋白的糖蛋白通过切下凝胶切片、胰酶消化和MALDI-TOF MS分析来鉴定。此外，这些糖蛋白上的糖链被释放，用吡啶胺标记，并通过层析分离和MALDI-TOF MS鉴定。初步结果表明，DBA表位位于N-连接的糖链上，因为它们对N-糖链的释放敏感。最终，我们将回答DBA多糖活性在分化后如何消失的问题。 我们正在鉴定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. We are identifying the glycoproteins that express this glycan and their function.