Description
List of abbreviations Preface Introduction Chapter 1 The World of Carbohydrates Chapter 2 Basic Knowledge of Glycobiology 2.1 Monosaccharides 2.1.1 The Molecular Structure 2.1.2 D/L-forms of Monosaccharides 2.1.3 Specific Monosaccharides 2.2 Oligosaccharides 2.3 Polysaccharides 2.4 Glycoconjugates 2.4.1 Glycoproteins 2.4.1.1 N-glycans 2.4.1.2 O-glycans 2.4.1.3 Blood Group Determinants 2.4.1.4 Different Types of Glycosylation 2.4.1.5 Glycosides 2.4.2 Proteoglycans and Glycosaminoglycans 2.4.3 Glycolipids 2.4.4 Glycosylphosphatidyl-inositol Membrane Anchors 2.5 Intrinsic Complexity of Carbohydrates 2.6 Working in a Laboratory Chapter 3 Carbohydrates involved in Diseases Chapter 4 Detection of Carbohydrate by Colorimetric Methods 4.1 Introduction 4.2 Preparing Standard Sugar Solutions for Calibration Curves 4.3 General tests 4.3.1 Detection of Carbohydrate by a Spot Test 4.3.2 Phenol-Sulfuric Acid Assay 4.3.3 Anthrone Test 4.3.4 Molisch’s Test 4.3.5 Fehling’s Test 4.3.6 Benedict’s Test 4.3.7 Barfoed’s Test 4.4 Specific Tests 4.4.1 Seliwanoff’s Test for Ketoses 4.4.2 Bial’s Test for Pentoses 4.4.3 Colorimetric Analysis of (N-acetyl) Aminosugars 4.4.4 Colorimetric Analysis of Uronic Acids 4.4.5 Colorimetric Analysis of Sialic Acids 4.4.6 Iodine Test for Starch Chapter 5. Analytical Techniques to Study Carbohydrates 5.1 Structural Parameters 5.2 Glycosidic-bond Cleavage techniques 5.2.1 Sulfuric Acid Hydrolysis 5.2.2 Hydrochloric Acid Hydrolysis 5.2.3 Trifluoroacetic Acid Hydrolysis 5.2.4 Methanolysis 5.2.5 Formolysis 5.2.6 Acetolysis 5.2.7 Specific Degradations 5.3 Common Separation Techniques for Mono/Oligosaccharides 5.3.1 Thin-Layer Chromatography 5.3.2 Size-Exclusion Chromatography 5.3.3 High-Performance Liquid Chromatography 5.3.4 Porous Graphitized Carbon Chromatography 5.3.5 Anion/Cation-Exchange Chromatography 5.3.6 High-pH Anion-Exchange Chromatography 5.3.7 Affinity Chromatography 5.4 Detection Techniques for Liquid Chromatography 5.4.1 Detectors 5.4.2 Labeling of Glycans for Detection 5.4.2.1 Labeling by Reductive Amination 5.4.2.2 Labeling by Michael Addition with PMP 5.4.2.3 Permethylation of Glycans for Mass Spectrometry 5.5 Electrophoretic Separation of Carbohydrates 5.6 Gas-Liquid Chromatography Chapter 6 Monosaccharide Composition Analysis 6.1 Introduction 6.2 Gas-chromatographic Monosaccharide Analysis using TMS Methyl Glycosides 6.3 Gas-chromatographic Monosaccharide Analysis using Alditol Acetates 6.4 Determination of D/L Configuration of Monosaccharides 6.5 Monosaccharide Analysis by HPLC Methods 6.5.1 High-pH Anion-Exchange Chromatography 6.5.2 Ultra-High-Performance Liquid Chromatography 6.6 Determination of Carbohydrate Linkages by Methylation Analysis 6.6.1 Chemical Aspects 6.6.2 GLC-EI Mass Spectrometry of PMAAs 6.6.3 Glycosidic Linkage Determination by HPLC-MS/MS Chapter 7 Carbohydrate Analysis of Glycoconjugates 7.1 Introduction 7.2 Analysis of Glycoproteins 7.2.1 A Typical Approach to the Analysis of a Glycoprotein 7.2.2 Chemical Release of N- and O-linked Glycans 7.2.3 Enzymatic Release of N-linked Glycans 7.2.3.1 Isolation of N-glycans and de-N-glycosylated protein (containing O-glycans) 7.2.4 Release of O-glycans by Reductive -elimination 7.3 Analysis of Glycopeptides 7.3.1 Preparation of Glycopeptides from Glycoproteins 7.3.2 Separation of Glycopeptides 7.4 Analysis of Proteoglycans and their Glycosaminoglycans 7.5 Analysis of Glycolipids 7.5.1 General Aspects 7.5.2 Thin-Layer Chromatography of Glycolipids 7.5.3 Carbohydrates of Glyco(sphingo)lipids 7.5.4 Carbohydrates of Glycosylphosphatidylinositol (GPI) Anchors 7.6 Analysis of Polysaccharides Chapter 8 Structural Characterization of Released Glycans 8.1 Introduction 8.2 Analysis of N-glycans 8.2.1 Fractionation of N-glycans 8.2.2 Preparation of Glucose Oligomers Standard (Dextran-ladder) 8.2.3 Enzymatic Sequence Analysis of N-glycans 8.3 Analysis of O-glycans 8.3.1 General Aspects 8.3.2 Mucin Glycoproteins 8.3.3 Fractionation of O-glycans Chapter 9 Analysis of Sialic Acids 9.1 General Aspects 9.2 Characterization of Sialic Acid Residues 9.2.1 Colorimetric Determination 9.2.2 Gas-chromatographic Determination 9.2.3 Release of Sialic Acids by Mild Acid Hydrolysis 9.2.4 Analysis of DMB-labeled Sialic Acid 9.2.5 Release of Sialic Acids by Enzymatic Digestion 9.3 Mass Spectrometry of Sialic Acids 9.4 1H NMR Spectroscopy of Sialic Acids Chapter 10 Carbohydrate Microarray Technology Chapter 11 Analysis of Carbohydrates by Mass Spectrometry 11.1 General Aspects 11.2 Electron Impact Mass Spectrometry (EI-MS) 11.3 Mass Spectrometry of Glycans 11.3.1 General 11.3.2 Matrix-Assisted Laser Desorption/ionization-Time of Flight mass spectrometry (MALDI-TOF-MS) 11.3.3 Electrospray Ionization-Mass Spectrometry (ESI-MS) 11.3.4 Tandem Mass Spectrometry 11.3.5 Mass Fragmentation of Oligosaccharides Chapter 12 Analysis of Carbohydrates by Nuclear Magnetic Resonance Spectroscopy 12.1 Introduction 12.2 Principle of NMR Spectroscopy 12.3 NMR Spectroscopy of Carbohydrates 12.3.1 Sample Preparation and Measuring 12.3.2 1H NMR Structural-Reporter-Group Concept 12.3.3 13C NMR Spectroscopy 12.4 NMR Spectroscopy for Glycans 12.4.1 Two-dimensional NMR Spectroscopy 12.4.2 Spectral Interpretation and Assignments 12.5 NMR Spectroscopy of Polysaccharides 12.5.1 Summarized NMR Measuring Procedure Chapter 13 Glycobioinformatics Chapter 14 Concluding Remarks Appendix A EI-MS of partially methylated alditol acetates (PMAAs) Appendix B MALDI-TOF-MS of permethylated N-glycans Appendix C ES-MS [M-H]- signals of some O-glycans (as alditols) Glossary of terms ACKNOWLEDGMENTS DISCLAIMER REFERENCES Subject Index
