A novel class of glycoconjugates has been identified and structurally characterized. These glycoconjugates contain fucose O-linked to amino acid, e.g. FL3a, fucosylAlpha1 (e.g. FL4a, glucosylBeta(1-3) fucosylAlpha1 greater than threonine). These glycoconjugates are of interest not only because of their novel structures but because the metabolism of these amino acid fucosides is altered in a variety of oncogenically-transformed cells. In addition, the metabolism of these compounds is regulated in normal cells as a function of cell density. The novel structures of the amino acid fucosides raises the possibility of proteins with O-linked fucose. There is preliminary evidence for the presence of related and possibly precursor protein(s) with O-linked fucose and O-linked glucosyl fucose. Hence, the major objectives of this proposal are to characterize glycoproteins with O-linked fucose contrituents with regard to size, charge, metabolism, subcellular localization; to pinpoint the biochemical basis of the altered metabolism of compounds containing O-linked fucose constituents in transformed cells and to examine the regulation of the metabolism of these compounds as a function of cell density. These studies will involve gel filtration, lectin affinity chromatography, and polyacrylamide gel electrophoresis. Metabolic labeling with (3H)-fucose will involve pulse, pulse/chase, long term labeling and long term labeling/chase experiments. Cell-free studies are anticipated to focus on the reactions of interest. Thus, the combined studies should define these novel fucoconjugates with respect to structure, metabolism, subcellular localization and in turn should define the basis for their alterations in cancer cells.
NIH ExPorter
NIH ExPorter