The investigation of the structural and functional roles of glycoprotein oligosaccharides by Fourier-transform mass spectrometry (FTMS)

The first chapter briefly outlines the application of matrix-assisted laser desorption/ionization Fourier-transform mass spectrometry (MALDI-FTMS) on the analysis of the O-linked oligosaccharide mixture from the egg-jelly glycoproteins of Xenopus laevis, a South-African frog. Methods for the releasing by alkaline reductive elimination and purification by P2-Gel and amine columns were described. The suitability of MALDI-FTMS analysis of the mixture was validated.; The second chapter describes the catalog-library approach (CLA) for the complete structural determination of neutral O-linked oligosaccharides including sequence, linkage, and even stereochemistry in the picomole level. By combining motifs found in the unknown, the new structure could be elucidated.; Chapter three and four summarize the structural analysis of neutral and anionic O-linked oligosaccharide alditol mixtures from the egg jelly glycoprotein of X. laevis respectively. Various mass spectrometry techniques including alkali metal doping, exact mass calculation, CID, and the catalog-library approach (CLA) for the determination of oligosaccharide structures are discussed in detail.; Chapter five deals with the construction of a bio-affinity MALDI probe in the determination of biologically active oligosaccharides to the cortical granule lectin (CGL). A host of other known lectins with their active oligosaccharides were first tested for the validity of the probe. Then, the CGL binding oligosaccharides were determined and their structures elucidated. Variations between terminal structures on binding and non-binding oligosaccharides provided some in sight into the mechanism of the lectin binding.; Chapter six describes a faster and easier method for the analysis of a complex mixture by combining open-channel capillary electrophoresis with mass spectrometry on a chip.{09}It tackled the important problem of the high through-put, or the lack of it, in the current age of “industrial biology.” The Rapid Open Channel Electrophoresis (ROCHE) device was fabricated and it produced some encouraging results.; Chapter seven pays tribute to the remarkable diversity in the oligosaccharide expression among members of the same species. A rudimentary separation by high performance liquid chromatography (HPLC) followed by MALDI-FTMS analysis provided the solid evidence of this diversity. This method is able to identify specific structures rapidly (minutes) with high sensitivity (picomoles or less) and may be important in the field of evolutionary biology.