Glycobiology

In the past decade, there has been an explosion of progress in understanding the roles of carbohydrates in biological systems. This explosive progress was made with the efforts in determining the roles of carbohydrates in immunology, neurobiology and many other disciplines, examining each unique system and employing new technology. This volume represents the first of three in the Methods in Enzymology series, including Glycomics (vol. 416) and Functional Glycomics (vol. 417), dedicated to disseminating information on methods in determining the biological roles of carbohydrates. These books are designed to provide an introduction of new methods to a large variety of readers who would like to participate in and contribute to the advancement of glycobiology. The methods covered include structural analysis of carbohydrates, biological and chemical synthesis of carbohydrates, expression and determination of ligands for carbohydrate-binding proteins, gene expression profiling including micro array, and generation of gene knockout mice and their phenotype analyses.

Section 1 - N-Glycan processing
Non-radioactive analysis of lipid-linked oligosaccharide compositions by fluorophore-assisted carbohydrate electrophoresis (FACE).
Identification of N-glycan-binding proteins for E3 ubiquitin ligases.
Familyi 47 á-mannosidases in N-glycan processing.
A Cytoplasmic Peptide: N-glycanase.

Section 2 - Structural Analysis
Glycomic profiling of cells and tissues by mass spectrometry – fingerprinting and sequencing methodologies.
Structural analysis of sialyl N-glycan using pyridylamination and chromatography followed by multistage tandem mass spectrometry.
Determination of glycosylation sites and disulfide bond structures using LC/ESI-MS/MS analysis.
Identification of O-GlcNAc sites on proteins.

Section 3 - Carbohydrate Synthesis and Antibiotics
Chemoenzymatic synthesis of glycan libraries.
Glycoconjugate vaccines against Haemophilus influenzae tybe b.
Assay of human gastric mucin as a natural antibiotic against Helicobacter pylori.
Molecular contacts between antibiotics and the 30S ribosomal particle.
Mechanism-based inhibitors to probe transitional states of glycoside hydrolases.
Regulating cell surface glycosylation with a small molecule switch.
Metabolic labeling of glycans with azido sugars for visualization and glycoproteomics.

Section 4 - Carbohydrate Ligand Specificity
Functional Proteomic Profiling of Glycan Processing Enzymes.
Oligosaccharide Microarrays to Map Interactions of Carbohydrates in Biological Systems.
Identification of Ligand Specificities for Glycan-binding Proteins Using Glycan Arrays.
High-throughput analysis of lectin-oligosaccharide interactions by automated frontal affinity chromatography.
Preparation of Neoglycolipids with Ring-Closed Cores via Chemoselective Oxime-Ligation for Microarray Analysis of Carbohydrate-Protein Interactions.
Development of a lectin microarray based on an evanescent-field fluorescence principle.