Lectin Methods and Protocols

ISBN-13: 9781617370243 / Angielski / Miękka / 2010 / 616 str.

Jonathan M. Rhodes; Jeremy D. Milton

Opis

Lectins have in the past been regarded by many scientists as curious proteins of uncertain structure and specificity that bind to carbohydrates of dubious significance themselves. All this is rapidly changing. The functional importance of glycosylation in cell-cell and cell-pathogen interactions, as well as intracellular events, has been recognized by the explosion of the science of glycobiology. This has been paralleled by the realization that lectins, once they have been well characterized, can be extremely useful tools for exam- ing structural changes in glycosylation and their functional consequences for human pathophysiology. Different lectins vary considerably in their degree of specificity. Some, such as wheatgerm agglutinin, have fairly broad specificity (for glucosamine or sialic acid), whereas others, such as Maackia amurensis, are specific not only for a single carbohydrate, but also for its linkage (2-3 linked sialic acid). Lectins with relatively broad specificity may be very useful as an adjunct to isolation or quantification of soluble glycoproteins, whereas lectins of known, and precise, specificity will be more useful for characterization of carbo- drate structure. We have included an appendix in Lectin Methods and Pro- cols that provides the known specificities of all lectins cited in the text.

Spis treści

Part I. Lectin Histochemistry and Cytochemistry. A. Light Microscopy. Light Microscopy: Overview and Basic Methods, Anthony J. Leathem and Susan A. Brooks. Avidin-Biotin Amplifications on Araldite-Embedded Sections, Robert W. Stoddart and Carolyn J. P. Jones. Lectin-Gold Histochemistry on Paraffin and Lowicryl K4M Sections using Biotin and Digoxigenin-Conjugated Lectins, Juergen Roth, Christian Zuber, Tetsutaro Sata, and Wei-ping Li. Use of Fluorochrome-Labeled Lectins in Light Microscopy, Udo Schumacher and Barry S. Mitchell. The Use of Lectins in Combination with Enzymatic Digestion for the Study of Glycoconjugates in Cartilage, Sibylle Hoedt-Schmidt. Applications of Lectin Histochemistry and Cytochemistry in Diagnosis and Prognosis, Timothy Richard Helliwell. B. Electron Microscopy. Electron Microscopy: Use of Lectin-Peroxidase Prior to Embedding, Adi Ellinger. Use of Lectin-Gold After Embedding, Reiner Herken and Berti Manshausen. Amplification of Lectin-Gold Histochemistry, Juan F. Madrid, Francisco Hernandez, and José Ballesta. Electron Microscopic Methods for the Demonstration of Lectin-Binding Sites in Cancer Cell Lines, Bruce S. Mitchell and Udo Schumacher. Part II. Use of Lectins for Structural Analysis of Oligosaccharide Chains. Application of Sequential Smith Degradation to Lectin Blots, Chi Kong Ching. Blot Analysis with Lectin for the Evaluation of Glycoproteins in Cultured Cells and Tissues, Christian Zuber, Wei-ping Li, and Jürgen Roth. Characterization of HIV gp120 Envelope Glycoprotein by Lectin Analysis, Gregers J. Gram and John-Erik Stig Hansen. Use of Lectins for Characterization of O-Linked Glycans of Herpes Simplex Virus Glycoproteins, Sigvard Olofsson and Anders Bolmstedt. Part III. Lectins for Detection of Altered Glycosylation of Circulating Glycoproteins. Use of Lectin for Detection of Agalactosyl IgG, Naoyuki Tsuchiya, Tamao Endo, Naohisa Kochibe, Koji Ito, and Akira Kobata. Lectins for Detection of Altered Glycosylation of CirculatingGlycoproteins: a-1 Antitrypsin, Yutaka Aoyagi and Hitoshi Asakura. a-Fetoprotein Using Lectin-Affinity Electrophoresis, Kazuhisa Taketa, Miao Liu, and Hiroko Taga. Use of Lectin-Affinity Electrophoresis for Quantification and Characterization of Glycoforms of a-I Acid Glycoprotein, Thorkild C. Bøg-Hansen. ABO(H) Blood Group Expression on Circulating Glycoproteins, Taei Matsui and Koiti Titani. Part IV. Use of Lectins in Quantification of Soluble Glycoproteins. Lectin/Antibody 'Sandwich' ELISA for Quantification of Circulating Mucin as a Diagnostic Test for Pancreatic Cancer, Neil Parker. Quantification of Intestinal Mucins, Jeremy D. Milton and Jonathan M. Rhodes. Part V. Lectins in Affinity Purification of Soluble Glycoproteins. Purification and Characterization of Human Serum and Secretory IgA1 and IgA2 Using Jacalin, Michael A. Kerr, Lesley M. Loomes, Brian C. Bonner, Amy B. Hutchings, and Bernard W. Senior. Use of Lectins in Affinity Purification of HIV and SIV Envelope Glycoproteins, Gustav Gilljam. T-Cell Receptor Purification, Kelly P. Kearse. Part VI. Lectins in Flow Cytometry. Use of Monomeric, Monovalent Lectin Derivatives for Flow Cytometric Analysis of Cell Surface Glucoconjugates, Hanae Kaku and Naoto Shibuya. Analysis of Subcellular Components by Fluorescent-Lectin Binding and Flow Cytometry, Rosa M. Guasch and José-Enrique O'Connor. Part VII. Lectins as Tools for Cell Purification/Purging. Lectins as Tools for Purification of Liver Endothelial Cells, Daniel E. Gomez and Unnur P. Thorgeirsson. The Use of Soybean Agglutinin (SBA) for Bone Marrow (BM) Purging and Hematopoietic Progenitor Cell Enrichment in Clinical Bone Marrow Transplantation, Arnon Nagler, Shoshana Morecki, and Shimon Slavin. Combined Lectin/Monoclonal Antibody Purging of Bone Marrow for Use in Conjunction with Autologous Bone Marrow Transplantation in the Treatment of Multiple Myeloma, Elizabeth G. H. Rhodes. Part VIII. Effects of Lectins on Mammalian Cells. Mechanisms and

Blurb

Lectin Methods and Protocols brings together a comprehensive collection of optimized methods for using lectins in biomedical research. The techniques-fine-tuned by the researchers who developed them-apply lectins to the analysis of carbohydrates, to the isolation and quantification of glycoproteins, and to living cells. Also included are step-by-step methods for the separation of human bone marrow cells for clinical use, for the assessment of the biological effects of plant lectins, and for the use of light and electron microscopy in analyzing carbohydrate structures. Taken together, the methods detailed here offer extremely useful tools for examining structural changes in glycosylation and their functional consequences for human pathophysiology

Lectin Methods and Protocols provides invaluable, easily reproducible techniques suitable for application to a wide range of important biomedical problems. The book will immediately become the standard practical resource and guide to lectin techniques for today's biomedical researchers.