GLYCOPROTEIN GLYCOMICS: EXPRESSION IN MOUSE ES CELLS & PROGENY

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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.