Lectins and Glycobiology

The intriguing complexity, precision, and regulation of the wide range of biologi­ cal processes is determined by intricate mechanisms of molecular recognition. Their nature is under intense scrutiny. In addition to the well-appreciated interac­ tion of proteins either with amino acid or nucleotide sequences, the investigation of their interplay with carbohydrate elements of cellular glycoconjugates current­ ly exerts increasing attraction. In the group of carbohydrate-binding proteins, lec­ tins are distinguished from antibodies or ligand-affecting enzymes, according to the most recent definition. The thorough analysis of their structure and function is considered as a focus to collect a critical mass of information for delineating details of a further array of biochemical processes with pivotal physiological im­ pact. Following an already century-long history of scientific description, reflected by subjectively chosen highlights (see the Brief History of Lectin Research at page VI), the excitement in glycobiological research that prevails today can easily be ex­ plained by our growing awareness of the multifarious significance of a sugar-code system of biological information. This present notion unmistakably has an im­ pact on lines of research in diverse disciplines like cell and molecular biology, histochemistry, or clinical sciences. It also prompts inherent practical questions such as how to obtain lectins, or how to employ them as instruments in various assay systems with the best possible results. Thus, this book is devoted intentionally to cover the techniques in different research fields that deal with lectins.

1 Lectins: Insights into the State of Knowledge by Literature Searches.- Chemical Synthesis of Lectin Ligands.- 2 Synthetic Ligands for Lectins.- 3 Sugar-Polyacrylamide Conjugates as Probes for Cell Lectins.- Purification and Characterization of Lectins.- 4 Isolation of Plant Lectins.- 5 A General Procedure for the Purification of Fimbrial Lectins from Escherichia coli.- 6 Isolation, Detection, and Ligand Binding Specificity of a Lectin Uniquely Found in Rat Kupffer Cells.- 7 Galaptin: Isolation, Electrophoretic Analysis, Peptide Preparation for Sequencing and Carbohydrate-Binding Site Specificity.- 8 Determination of Carbohydrate Specificity in Solid-Phase Assays.- 9 Analysis of Isoelectric Variants of Carbohydrate-Binding Protein 35.- 10 Chemical Modification of Lectins by Group-Specific Reagents.- Biophysical Methods for the Characterization of Lectin-Ligand Interactions.- 11 Study of Oligosaccharide-Lectin Interaction by Various Nuclear Magnetic Resonance (NMR) Techniques and Computational Methods.- 12 Lectin-Glycoconjugate Cross-Linking Interactions.- 13 Lectins as Analytical Probes to Define the Physical Characteristics of Binding Events.- Lectins as Tools for the Characterization of Glycoconjugates.- 14 Glycoprotein-Lectin-Immunosorbent Assay (GLIA).- 15 Use of Lectins in Quantification and Characterization of Soluble Glycoproteins.- 16 Lectins as Indicators of Disease-Associated Glycoforms.- 17 Viral Lectins for the Detection of 9-O-Acetylated Sialic Acid on Glycoproteins and Glycolipids.- 18 Affinoblotting Detection of the Glycoprotein Ligands of the Endogenous Cerebellar Soluble Lectin CSL and of Concanavalin A.- Determination of Expression of Lectins and Their Ligands.- 19 An Enzyme Immunoassay (EIA) for a Human Endogenous ?-Galactoside-Binding Lectin.- 20 Analysis of Lectin Expression by Immunoblotting.- 21 Application of Immuno- and Affinocytochemistry to the Detection of an Endogenous ?-Galactoside-Specific 14 kDa Lectin and Its Ligands.- Lectins and Neoglycoconjugates in Histochemical and Cytochemical Analysis.- 22 Tissue Lectins in Histopathology ― Markers in Search of Their Physiological Ligands.- 23 Lectin Cytochemistry Using Colloidal Gold Methodology.- 24 Neoglycoprotein-Gold Complexes as a Tool in the Study of Carbohydrate-Specific Binding Proteins.- 25 Lectins and Neoglycoproteins ― Attractive Molecules to Study Glycoconjugates and Accessible Sugar-Binding Sites of Lower Vertebrates Histochemically.- 26 In Situ Microdensitometry of Neoglycoprotein Staining: Detection of Tissue-Site Specific Patterns of Endogenous Lectin Expression.- 27 Lectin and Neoglycoprotein Binding to Cells.- 28 Use of Lectins for the Phenotypic Characterization of Cultured Cell Monolayers.- 29 The Use of Lectins in the Purification of Cellular Subpopulations.- 30 Lectin-Binding Properties of Extracellular Matrix in the Chick Oral Membrane.- 31 Biodistribution of Radioactive Lectins.- 32 Lectin Interactions with Intestinal Epithelial Cells and Their Potential for Drug Delivery.- 33 Neoglycoprotein-Liposome Conjugates for Studies of Membrane Lectins.- Lectins and Biosignaling.- 34 Effects of Lectins on Adenylylation and Phosphorylation of Plasma Membrane Proteins.- 35 Lectin-Induced Alterations in the Level of Phospholipids, Inositol Phosphates, and Phosphoproteins.- 36 Measurement of Intracellular Calcium Levels by Flow Cytometry Following Treatment of Murine Macrophage/Monocytes with Mistletoe Lectin.- 37 Measurement of Lectin-Induced Superoxide Release from Human Neutrophils.- 38 Isolation of Lymphocytes and Accessory Cells for Assays of Mitogenicity.- 39 Determination of Lectin-Mediated Augmentation of Natural Killer Cell-Mediated Cytotoxicity.- 40 Effects of Pseudomonas aeruginosa PA-I and PA-II Lectins on Tumoral Cells.- 41 Determination of Lectin-Dependent Alterations of Cellular Parameters by Immunophenotyping During Adjuvant Lectin Application.- 42 Lectin-Dependent Alteration in Availability of Nutrients in Serum and Erythrocytes.- Lectins and Cell Adhesion.- 43 Bacterial Adhesion to Immobilized Carbohydrates.- 44 Quantitative Micro-Adhesion Assay on Polystyrene Matrices.- 45 Lectin-Mediated Binding Activities of Bradyrhizobium japonicum.- 46 Identifying a Role for Carbohydrate Recognition in Regulating Neuronal Architecture.- 47 Identifying Cell-Cell Adhesion-Inhibitory Antibodies to Cell Surface Proteins Using Divalent Primary and Monovalent Secondary Antibodies ― a Method Developed Using Cellular Slime Molds.- 48 A Modified Capillary Method for Measuring Bacterial Chemotaxis.- Molecular Genetics of Lectins.- 49 Cloning Lectin Genes.- 50 Cell-Free Synthesis of Lectins.- 51 cDNA Cloning and Expression of Plant Lectins from the Legume Family.- 52 Production of Intact Recombinant Lectins in Escherichia coli.- 53 Construction of Mutant Lectin Genes by Oligonucleotide-Directed Mutagenesis and Their Expression in Escherichia coli.- 54 Mammalian Lectin as Transforming Growth Factor.- 55 Identification and Expression of a Protozoan Invasion Factor: Penetrin, the Heparin-Binding Protein from Trypanosoma cruzi.