NMR METHODOLOGY FOR CHARACTERIZING CARBOHYDRATE-PROTEIN INTERACTIONS (TB1)

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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. This project is part of a more general integrated technology project directed at the structural characterization of glycoaminoglycan-protein complexes. The formation of carbohydrate-protein complexes, in general, is important in a variety of processes involving the interaction of a cell with its environment. These processes include physiological processes, such as cell differentiation, cell aggregation, and cell signaling; they also include disease processes, such as viral infection, malignancy, and unwanted inflammation. The development of drugs to moderate natural processes and inhibit disease processes begins with accurate structural models for oligosaccharide-protein interactions. These models have been difficult to get by traditional Nuclear Magnetic Resonance (NMR) approaches because of the dearth of short-distance NOE contacts in oligosaccharide-protein complexes. We are developing a number of experiments and analysis tools that allow retrieval of structural information when traditional NMR approaches fail. This particular project combines new isotopic labeling strategies developed in the Moremen laboratory with new methods of assignment, and new types of NMR-observable structural data. The target systems include both glycosylated proteins that are difficult to express in forms amenable to use of NMR strategies that are dependent on uniform labeling with multiple NMR-active isotopes, and high order aggregates including chemokine-glycosaminoglycan (GAG) aggregates, where resolution and sensitivity become a problem.