Renato V. Iozzo (ed.)

Proteoglycan Protocols

Herausgegeben:Iozzo, Renato V.

• Gebundenes Buch

Produktbeschreibung

Proteoglycans are some of the most elaborate macromolecules of mammalian and lower organisms. The covalent attachment of at least five types of glycosami- glycan side chains to more than forty individual protein cores makes these molecules quite complex and endows them with a multitude of biological functions. Proteoglycan Protocols offers a comprehensive and up-to-date collection of prepa- tive and analytical methods for the in-depth analysis of proteoglycans. Featuring st- by-step detailed 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, to generate strategies for prokaryotic and eukaryotic expression, to utilize methods for the suppression of specific proteoglycan gene expression and for the detection of mutant cells and degradation products, and to study specific interactions between proteoglycans and extracellular matrix proteins as well as growth factors and their receptors. The readers will find concise, yet comprehensive techniques carefully drafted by leading experts in the field. Each chapter commences with a general Introduction, followed by a detailed Materials section, and an easy-to-follow Methods section. An asset of each chapter is the extensive notation that includes troubleshooting tips and practical considerations that are often lacking in formal methodology papers. The reader will find this section most valuable because it is clearly provided by experienced scientists who have first-hand knowledge of the techniques they outline. In addition, most of the chapters are well illustrated with examples of typical data generated with each method.

Produktdetails

• Methods in Molecular Biology 171
• Verlag: Humana / Humana Press / Springer, Berlin
• Artikelnr. des Verlages: 978-0-89603-759-5
• 2001
• Seitenzahl: 580
• Erscheinungstermin: 1. Juni 2001
• Englisch
• Abmessung: 235mm x 157mm x 36mm
• Gewicht: 1016g
• ISBN-13: 9780896037595
• ISBN-10: 0896037592
• Artikelnr.: 22447015

Inhaltsangabe

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.

Rezensionen

"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 clichés 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