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Full Description
Glycoconjugates such as glycoproteins and glycolipids play important roles in cell-cell interaction events, including development, differentiation, m- phogenesis, fertilization, inflammation, and metastasis. A number of reports have documented the association of unique oligosaccharide sequences to p- tein targeting and folding, and in mechanisms of infection, inflammation, and immunity. For glycoproteins, these glycan appendages are the result of extensive co- or post-translational modifications of the nascent proteins in the endoplasmic reticulum and in the Golgi apparatus. Although nucleic acids and proteins are copied from a template in a repeated series of identical steps using the same enzymes, complex carbohydrates are formed by the sequential actions of cellular glycosyltransferases that specifically recognize unique s- strates. The molecular biology of these transferases and other carbohydra- modifying enzymes is providing important insights on oligosaccharide recognition events.
While it is acknowledged that the definition of the protein complement of cells and tissues (the so-called proteome) remains an enormous task in this postgenomic era, the characterization of all glycans produced by individual organisms (referred to as the glycome) presents an equally imp- tant challenge. This task is further complicated by the fact that oligosacc- rides cannot presently be cloned. These complex carbohydrates exist in a staggering diversity of structures, linkages, and branching, thus providing an exquisite molecular repertoire for cellular interactions.
Contents
Structural and Functional Diversity of Glycoconjugates.- Sample Preparation Methods.- Chemical and Enzymatic Release of Glycans from Glycoproteins.- Derivatization of Carbohydrates.- Preparation of Oligosaccharides from Sulfated Glycosaminoglycans Using Bacterial Enzymes.- Analysis of Mono- and Oligosaccharides.- Monosaccharide Analysis by Capillary Electrophoresis.- Separation of Oligosaccharides by Capillary Electrophoresis Using Buffer Modifiers.- Characterization of Oligosaccharides from Starch, Dextran, Cellulose, and Glycoproteins by Capillary Electrophoresis.- Affinity Capillary Electrophoresis of Oligosaccharides.- Characterization of Glycosaminoglycans by Capillary Electrophoresis.- Analysis of Glycoconjugates and the Component Monosaccharides.- Analysis of 2-Aminoacridone-Derivatized Complex Oligosaccharides Using Micellar Electrokinetic Capillary Chromatography and Laser-Induced Fluorescence Detection.- Separation of Protein Glycoforms by Capillary Electrophoresis.- Separation of Glycoproteins by Capillary Isoelectric Focusing.- Glycoprotein Analysis by Capillary Zone Electrophoresis-Electrospray Mass Spectrometry.- Analysis of Bacterial Glycolipids by Capillary Electrophoresis-Electrospray Mass Spectrometry.- Specific Applications.- Capillary Electrophoresis as an Assay Method for Monitoring Glycosyltransferase Activity.- Determination of Association Constant of Carbohydrate-Protein Interaction.
