Overview

Renato Iozzo and an authoritative team of investigators present for the first time a comprehensive and up-to-date collection of readily reproducible preparative and analytical methods for the in-depth analysis of these important compounds. Featuring step-by-step protocols, this book will enable both novice and experienced researchers to isolate intact proteoglycans from tissues and cultured cells, to establish the composition of their carbohydrate moieties, and to generate strategies for prokaryotic and eukaryotic expression. There are also detailed techniques for the suppression of specific proteoglycan genes, for the detection of mutant cells and their degradation products, and for studying specific interactions between proteoglycans and extracellular matrix proteins and with growth factors and their receptors.

Full Product Details

Author:   Renato V. Iozzo
Publisher:   Humana Press Inc.
Imprint:   Humana Press Inc.
Edition:   2001 ed.
Volume:   171
Dimensions:   Width: 15.50cm , Height: 3.60cm , Length: 23.50cm
Weight:   2.240kg
ISBN:  

9780896037595

ISBN 10:   0896037592
Pages:   558
Publication Date:   01 June 2001
Audience:   College/higher education ,  Professional and scholarly ,  Undergraduate ,  Professional & Vocational
Format:   Hardback
Publisher's Status:   Active
Availability:   In Print  
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Table of Contents

Isolation and Purification.- Isolation of Proteoglycans from Cell Cultures and Tissues.- Isolation of Proteoglycans from Tendon.- Purification of Proteoglycans from Mineralized Tissues.- Purification of Perlecan from Endothelial Cells.- Isolation and Characterization of Nervous Tissue Proteoglycans.- Analysis of Proteoglycans and Glycosaminoglycans from Drosophila.- Cartilage and Smooth Muscle Cell Proteoglycans Detected by Affinity Blotting Using Biotinylated Hyaluronan.- Preparation of Proteoglycans for N-Terminal and Internal Amino Acid Sequence Analysis.- High-Specific-Activity 35S-Labeled Heparan Sulfate Prepared from Cultured Cells.- Selective Detection of Sulfotransferase Isoforms by the Ligand Affinity-Conversion Approach.- Particulate and Soluble Glycosaminoglycan-Synthesizing Enzymes.- Disaccharide Composition of Hyaluronan and Chondroitin/Dermatan Sulfate.- Integral Glycan Sequencing of Heparan Sulfate and Heparin Saccharides.- Analytical and Preparative Strong Anion-Exchange HPLC of Heparan Sulfate and Heparin Saccharides.- Proteoglycans Analyzed by Composite Gel Electrophoresis and Immunoblotting.- Quantitation of Proteoglycans in Biological Fluids Using Alcian Blue.- Cellulose Acetate Electrophoresis of Glycosaminoglycans.- Disaccharide Composition in Glycosaminoglycans/Proteoglycans Analyzed by Capillary Zone Electrophoresis.- Intact and Oligomeric Glycosaminoglycans Analyzed with Capillary Electrophoresis.- Expression, Detection, and Degradation.- Recombinant Expression of Proteoglycans in Mammalian Cells.- Purification of Recombinant Human Decorin and Its Subdomains.- Prokaryotic Expression of Proteoglycans.- Proteoglycan Gene Targeting in Somatic Cells.- Constitutive and Inducible Antisense Expression.- Cell-Mediated Transfer of Proteoglycan Genes.- Morphological Evaluation of Proteoglycans in Cells and Tissues.- Expression and Characterization of Engineered Proteoglycans.- Intracellular Localization of Engineered Proteoglycans.- Selection of Glycosaminoglycan-Deficient Mutants.- Xyloside Priming of Glycosaminoglycan Biosynthesis and Inhibition of Proteoglycan Assembly.- Inhibition of Heparan Sulfate Synthesis by Chlorate.- Detection of Proteoglycan Core Proteins with Glycosaminoglycan Lyases and Antibodies.- Proteoglycan Core Proteins and Catabolic Fragments Present in Tissues and Fluids.- Degradation of Heparan Sulfate by Nitrous Acid.- Degradation of Heparan Sulfate with Heparin Lyases.- Degradation of Chondroitin Sulfate and Dermatan Sulfate with Chondroitin Lyases.- In Vitro Assay for Hyaluronan Synthase.- Hyaluronidase Activity and Hyaluronidase Inhibitors.- Detecting Hyaluronidase and Hyaluronidase Inhibitors.- Interacations.- Affinity Coelectrophoresis of Proteoglycan-Protein Complexes.- Binding Constant Measurements for Inhibitors of Growth Factor Binding to Heparan Sulfate Proteoglycans.- Interaction of Proteoglycans with Receptor Tyrosine Kinases.- Measuring Single-Cell Cytosolic Ca2+ Concentration in Response to Proteoglycans.- Serum Amyloid A Peptide Interactions with Glycosaminoglycans.- Interactions of Lipoproteins with Proteoglycans.- Hyaluronan and Hyaluronan-Binding Proteins Probes for Specific Detection.- Novel Confocal-FRAP Analysis of Carbohydrate-Protein Interactions Within the Extracellular Matrix.- Regulatory Roles of Syndecans in Cell Adhesion and Invasion.- Optical Biosensor Techniques to Analyze Protein-Polysaccharide Interactions.- Phage Display Technology to Obtain Antiheparan Sulfate Antibodies.- Tissue-Specific Binding by FGF and FGF Receptors to Endogenous Heparan Sulfates.

Reviews

The book is strategically subdivide into 3 sections (isolation and purification; expression, detection, and degradation; interactions) with a total of 51 chapters ( 19, 20, and 12 per subheading). Their contents dispel the cliches about work with sticky glycosaminoglycans being tedious, unrewarding, and mostly for the sake of old-fashioned systematics. I expect even experts in certain areas to benefit from the whole book, whose rightful place will be on the bench, not on the shelf. - ChemBioChem This book will appeal to both those new to the subject and experts in the field as the protocols outlined are both wide ranging, comprehensive, and easy to follow. strongly recommend it to those new to proteoglycan research, and would regard it as an essential addition to any proteoglycan research laboratory library. - Molecular Biotechnology ...this book will be a very useful tool for many specialists working in the fields of glycobiology, biotechnology, biochemistry, and experimental medicine. Detailed description of the methods employed for various aspects of proteoglycan analysis and comprehensive bibliography accompanying each chapter make this book especially valuable for a large audience of the mentioned specialists. Biochemistry

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