Glycoprotein Analysis in Biomedicine

As the editorof Glycoprotein Analysis in BiornedidneI have tried to bring together a comprehensive range ofdetailed protocols that will rendercarbohydratebiochemistryaccessible to all, andnotjust to the dedicated few. I wish to thank the authors, all ofwhom have striven significantly to realize this aim. There is now a growing interest in glycoprotein analysis from researchers in many disciplineswho realize thatprotein glycosylation can no longer be ignored. Oligosaccharide chains of glycoproteins form a diverse arrayofmolecules with an inherentlylarge contentof chemical information that is functional in biological recognition. In addition to theirspecificinteractionswithantibodiesandcarbohydrate binding proteins, the carbohydrate moieties of glycoproteins have profoundly important effects on the conformations, stability, antige nicity, and function ofthe protein components. Some specialized techniques, Le., NMRand MS, are introduced atthebeginningofthebooksince thesearecapableofdefiningexactly the diversityofoligosaccharide sequences,and are used to detect the growing number ofnew structures as well as to indentiry, in new sce narios, those previously documented. Together with the chromato graphic methods of purification and analysis discussed in Chapters 1-12, there now exists a powerful armamentarium for glycoprotein characterization. The more chemical techniques are documented in detail in the earliest chapters, whereas the enzymatic methods of releasing oligosaccharidesfrom protein areintroducedlater. Analysis ofglycoproteinsfrom mammalian, parasitic,bacterial,andviralorigins are each discussed.

Résumé
This is a collection of detailed protocols designed to make carbohydrate biochemistry accessible to a wider audience.

Contenu
A General Strategy for Glycoprotein Oligosaccharide Analysis.- Analysis of Asparagine-Linked Oligosaccharides by Sequential Lectin-Affinity Chromatography.- Sequence and Linkage Analysis of N-and O-Linked Glycans by Fast Atom Bombardment Mass Spectrometry.- Gas Chromatography and Gas Chromatography-Mass Spectrometry of Glycoprotein Oligosaccharides.- Microscale Sequencing of N-Linked Oligosaccharides of Glycoproteins Using Hydrazinolysis, Bio-Gel P-4, and Sequential Exoglycosidase Digestion.- Analysis of Sugar Chains by Pyridylamination.- Analysis of Carbohydrates in Glycoproteins by High-Performance Liquid Chromatography and High-Performance Capillary Electrophoresis.- The Microanalysis of Glycosyl-Phosphatidylinositol Glycans.- Analysis of Bacterial Glycoproteins.- Isolation and Fractionation of Glycoprotein Glycans in Small Amounts.- High-Mannose Chains of Mammalian Glycoproteins.- Release of O-Linked Glycoprotein Glycans by Endo-?-N-Acetylgalactosaminidase.- Immunological Detection of Glycoproteins on Blots Based on Labeling with Digoxigenin.- Glycosyltransferases as Tools in Cell Biological Studies.- Purification of the EGF Receptor for Oligosaccharide Studies.- Analysis of Sulfated Polysaccharide Conjugates.- Secreted Mucus Glycoproteins in Cell and Organ Culture.- Preparation of Polyclonal Antibodies to Native and Modified Mucin Antigens.- The Use of Immunoperoxidase to Characterize Gastrointestinal Mucin Epitopes.- The Application of Lectins to the Study of Mucosal Glycoproteins.- Detection of Tumor-Associated Expression of Carbohydrate-Binding Proteins (Lectins).- Scanning Tunneling Microscopy of Biopolymers.