ANALYTICAL TECHNIQUES FOR STRUCTURAL ANALYSIS

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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. Participants will learn basic techniques for the structural characterization of glycosaminoglycans (GAGs), which will include depolymerization of the GAG polysaccharides to its disaccharide constituents using appropriate enzymes. The disaccharides will be separated by ion exchange chromatography and the products will be detected by U.V. The disaccharides will also be derivatized with 2-cyanoacetamide and detected with fluorescent detection. In a separate experiment, the glycosaminoglycans will be hydrolyzed by nitrous acid and the products will be analyzed. Mass spectrometry techniques for di- and oligosaccharides derived from glycosaminoglycans will also be covered, as well as sulfate and polysaccharide dye binding assays. Experience with basic biochemical techniques is a prerequisite for participation.