PURIFICATION OF HIGH MANNOSE OLIGOSACCHARIDES

THE LONG-TERM OBJECTIVES OF THIS PROPOSAL ARE TO PREPARE PURE OLIGOSACCHARIDES FROM GLYCOPROTEINS POSSESSING ASPARAGINE-LINKED CARBOHYDRATE CHAINS AND TO USE THE OLIGOSACCHARIDES AND THEIR DERIVATIVES TO INVESTIGATE THE STRUCTURE, BIOSYNTHESIS AND FUNCTION OF THE GLYCAN CHAINS OF GLYCOPROTEINS. THE SPECIFIC AIMS OF THIS PROPOSAL ARE TOPURIFY AND CHARACTERIZE HIGH MANNOSE OLIGOSACCHARIDES FROM SOYBEAN PROTEINS. SOYBEAN AGGLUTININ AND 7S SOYBEAN PROTEINWILL BE PURIFIED, THEIR CARBOHYDRATE CHAINS RELEASED BY ENDO H AND N-GLYCANASE(R) DIGESTION, AND THE OLIGOSACCHARIDES ISOLATED AFTER CHROMATOGRAPHIC SEPARATION 2-AMINOPYRIDYL DERIVATIVES. THE OLIGOSACCHARIDES WILL BE CHARACTERIZED BY HPLC ANALYSIS, (1)H-MNR SPECTROSCOPY, METHYLATION ANALYSIS AND FAB MASS SPECTROSCOPY. THE PURIFIED OLIGOSACCHARIDES WILL BE SOLD TO THE RESEARCH COMMUNITY AS PART OF GENZYME'S GLYCOPROTEIN PRODUCT LINE. THE CARBOHYDRATES CAN BE USED AS STANDARDS FOR STRUCTURAL ANALYSIS, AS SUBSTRATES FOR ENZYMATIC REACTIONS, AND AS LIGANDS FOR EXAMINING THE ROLE OF THE GLYCANS IN THE BINDINGOF GROWTH FACTORS, HORMONES, TOXINS, VIRUSES, BACTERIA AND LECTINS TO CELLS. ONE POTENTIAL LARGE-SCALE APPLICATIONS FOR THE OLIGOSACCHARIDES IS AS A LIGAND TO TARGET THERAPEUTIC AGENTS TO MACROPHAGES.