Glycobiology Protocols

1. Front Matter

   Pages i-xviii

   PDF

2. Application of Fluorophore-Assisted Carbohydrate Electrophoresis for the Study of the Dolichol Pyrophosphate-Linked Oligosaccharides Pathway in Cell Cultures and Animal Tissues

   • Ningguo Gao

   Pages 1-12

3. Partial Purification of Mannosylphosphorylundecaprenol Synthase From Micrococcus luteus

   • Jeffrey S. Rush, Charles J. Waechter

   Pages 13-30

4. Assay of Dolichyl-Phospho-Mannose Synthase Reconstituted in a Lipid Matrix

   • John S. Schutzbach

   Pages 31-41

5. O-Mannosylation in Mammalian Cells

   • Tamao Endo, Hiroshi Manya

   Pages 43-56

6. Methods for Analysis of Unusual Forms of O-Glycosylation

   • Aleksandra Nita-Lazar, Robert S. Haltiwanger

   Pages 57-68

7. Structural Analysis of Naturally Occurring Sialic Acids

   • Johannis P. Kamerling, Gerrit J. Gerwig

   Pages 69-91

8. Nonradioactive Trans-Sialidase Screening Assay

   • Silke Schrader, Roland Schauer

   Pages 93-107

9. Structural Determination of O-Glycans by Tandem Mass Spectrometry

   • Catherine Robbe, Jean-Claude Michalski, Calliope Capon

   Pages 109-123

10. Detailed Structural Analysis of N-Glycans Released From Glycoproteins in SDS-PAGE Gel Bands Using HPLC Combined With Exoglycosidase Array Digestions

    • Louise Royle, Catherine M. Radcliffe, Raymond A. Dwek, Pauline M. Rudd

    Pages 125-143

11. Molecular Modeling of Glycosyltransferases

    • Anne Imberty, Michaela Wimmerová, Jaroslav Koča, Christelle Breton

    Pages 145-156

12. β-Galactoside α2,6-Sialyltransferase and the Sialyl α2,6-Galactosyl-Linkage in Tissues and Cell Lines

    • Fabio Dall’Olio, Nadia Malagolini, Mariella Chiricolo

    Pages 157-170

13. Visualizing Intracellular Distribution and Activity of Core2 β(1,6)N-Acetylglucosaminyltransferase-I in Living Cells

    • Maëlle Prorok-Hamon, Sylvie Mathieu, Assou El-Battari

    Pages 171-186

14. Gene Expression Analysis of Glycosylation-Related Genes by Real-Time Polymerase Chain Reaction

    • Juan J. García-Vallejo, Sonja I. Gringhuis, Willem van Dijk, Irma van Die

    Pages 187-209

15. Analysis of the Glycodynamics of Primary Osteoblasts and Bone Cancer Cells

    • Inka Brockhausen, Xiaojing Yang, Mark Harrison

    Pages 211-236

16. Micromethods for the Characterization of Lipid A-Core and O-Antigen Lipopolysaccharide

    • Cristina L. Marolda, Piya Lahiry, Enrique Vinés, Soledad Saldías, Miguel A. Valvano

    Pages 237-252

17. Assay for a Galactosyltransferase Involved in the Assembly of the O7-Antigen Repeat Unit of Escherichia coli

    • Inka Brockhausen, Walter A. Szarek, John G. Riley, Jason Z. Vlahakis

    Pages 253-265

18. High-Throughput Quantitation of Metabolically Labeled Anionic Glycoconjugates by Scintillation Proximity Assay Utilizing Binding to Cationic Dyes

    • Karen J. Rees-Milton, Tassos P. Anastassiades

    Pages 267-275

19. Quantitative Analysis of Mucins in Mucosal Secretions Using Indirect Enzyme-Linked Immunosorbent Assay

    • Pablo Argüeso, Ilene K. Gipson

    Pages 277-288

20. Molecular Dissection of the Mouse Zona Pellucida

    • Frederick W. K. Kan

    Pages 289-304

Glycobiology involves studies of complex carbohydrates and posttrans- tional modifications of proteins, and has become an important interdiscip- nary field encompassing chemistry, biochemistry, biology, physiology, and pathology. Although initial research was directed toward elucidation of the different carbohydrate structures and the enzymes synthesizing them, the field has now moved toward identifying the functions of carbohydrates. The pro- cols described in Glycobiology Protocols form a solid basis for investigations of glycan functions in health and disease. The cloning of many of the genes participating in glycosylation processes has helped to enhance our knowledge of how glycosylation is controlled, but has also added another dimension of complexity to the great heterogeneous variety of the structures of the oligos- charides of glycoproteins, proteoglycans, and glycolipids. A family of similar enzyme proteins exists for each glycosylation step. Glycosyltransferases are extremely specific for both the nucleotide sugar donor and the acceptor s- strate, but many other factors control sugar transfer, including the locali- tion and topology of enzymes, cofactors, possible chaperone proteins, and the availability of sugar acceptor substrates. The analysis of the intracellular organization of glycosylation and of the factors controlling the activities of the participating enzymes in the cell are important areas that need more research efforts. Another challenge for future research is to understand the glycodynamics of a cell, that is, how the cell responds to stimuli leading to biological and pathological changes in terms of alterations in glycosylation, and how this affects the biology of the cell.

• Lipid
• Oligosaccharid
• Polysaccharid
• biochemistry
• enzymes
• gene expression
• physiology
• proteins

"...a very valuable handbook for a wide audience of specialists involved in studies of carbohydrates and carbohydrate-containing biopolymers; glycobiologists, biochemists, bioorganic chemists, and molecular and cell biologists." -Biochemistry

• Department of Medicine, Queen’s University, Kingston, Canada

  Inka Brockhausen

• Department of Biochemistry, Queen’s University, Kingston, Canada

  Inka Brockhausen

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