Proteoglycan Protocols

About this book

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.

Contents

I. Isolation and Purification Isolation of Proteoglycans from Cell Cultures and Tissues Masaki Yanagishita Isolation of Proteoglycans from Tendon Kathryn G. Vogel and Julie A. Peters Purification of Proteoglycans from Mineralized Tissues Neal S. Fedarko Purification of Perlecan from Endothelial Cells John Whitelock Isolation and Characterization of Nervous Tissue Proteoglycans Yu Yamaguchi Analysis of Proteoglycans and Glycosaminoglycans from Drosophila William D. Staatz, Hidenao Toyoda, Akiko Kinoshita-Toyoda, Kimberlly Chhor, and Scott B. Selleck Cartilage and Smooth Muscle Cell Proteoglycans Detected by Affinity Blotting Using Biotinylated Hyaluronan James Melrose Preparation of Proteoglycans for N-Terminal and Internal Amino Acid Sequence Analysis Peter J. Neame High Specific-Activity 35S-Labeled Heparan Sulfate Prepared from Cultured Cells Nicholas W. Shworak Selective Detection of Sulfotransferase Isoforms by the Ligand Affinity-Conversion Approach Nicholas W. Shworak Particulate and Soluble Glycosaminoglycan-Synthesizing Enzymes: Preparation Assay and Use Geetha Sugumaran and Jeremiah E. Silbert Disaccharide Composition of Hyaluronan and Chondroitin/Dermatan Sulfate: Analysis with Fluorophore-Assisted Carbohydrate Electrophoresis Anna H. K. Plaas, L. West, Ronald J. Midura, and Vincent C. Hascall Integral Glycan Sequencing of Heparan Sulfate and Heparin Saccharides Jeremy E. Turnbull Analytical and Preparative Strong Anion-Exchange HPLC of Heparan Sulfate and Heparin Saccharides Jeremy E. Turnbull Proteoglycans Analyzed by Composite Gel Electrophoresis and Immunoblotting George R. Dodge and Ralph Heimer Quantitation of Proteoglycans in Biological Fluids Using Alcian Blue Madeleine Karlsson and Sven Bjornsson Cellulose Acetate Electrophoresis of Glycosaminoglycans Yanusz Wegrowski and Francois-Xavier Maquart Disaccharide Composition in Glycosaminoglycans/Proteoglycans Analyzed by Capillary Zone Electrophoresis Nikos K. Karamanos and Anders Hjerpe Intact and Oligomeric Glycosaminoglycans Analyzed with Capillary Electrophoresis Nikos K. Karamanos and Anders Hjerpe II. Expression, Detection, and Degradation Recombinant Expression of Proteoglycans in Mammalian Cells: Utility and Advantages of the Vaccinia Virus/T7 Bacteriophage Hybrid Expression System David J. McQuillan, Neung-Seon Seo, Anne M. Hocking, and Camille I. McQuillan Purification of Recombinant Human Decorin and Its Subdomains Anna L. McBain and David M. Mann Prokaryotic Expression of Proteoglycans Alan D. Murdoch and Renato V. Iozzo Proteoglycan Gene Targeting in Somatic Cells Giancarlo Ghiselli and Renato V. Iozzo Constitutive and Inducible Antisense Expression Melissa Handler and Renato V. Iozzo Cell-Mediated Transfer of Proteoglycan Genes Jens W. Fischer, Michael G. Kinsella, David Hasenstab, Alexander W. Clowes, and Thomas N. Wight Morphological Evaluation of Proteoglycans in Cells and Tissues Stephanie L. Lara, Stephen P. Evanko, and Thomas N. Wight Expression and Characterization of Engineered Proteoglycans Wei Luo, Chunxia Guo, Jing Zheng, and Marvin L. Tanzer Intracellular Localization of Engineered Proteoglycans Tung-Ling L. Chen, Peiyin Wang, Seung S. Gwon, Nina W. Flay, and Barbara M. Vertel Selection of Glycosaminoglycan-Deficient Mutants Xiaomei Bai, Brett Crawford, and Jeffrey D. Esko Xyloside Priming of Glycosaminoglycan Biosynthesis and Inhibition of Proteoglycan Assembly Timothy A. Fritz and Jeffrey D. Esko Inhibition of Heparan Sulfate Synthesis by Chlorate H. Edward Conrad Detection of Proteoglycan Core Proteins with Glycosaminoglycan Lyases and Antibodies John R. Couchman and Pairath Tapanadechopone Proteoglycan Core Proteins and Catabolic Fragments Present in Tissues and Fluids John D. Sandy Degradation of Heparan Sulfate by Nitrous Acid H. Edward Conrad Degradation of Heparan Sulfate with Heparin Lyases Laurie A. LeBrun and Robert J. Linhardt Degradation of Chondroitin Sulfate and Dermatan Sulfate with Chondroitin Lyases Maria Jose Hernaiz and Robert J. Linhardt In Vitro Assay for Hyaluronan Synthase Andrew P. Spicer Hyaluronidase Activity and Hyaluronidase Inhibitors: Assay Using a Microtiter-Based System Susan Stair Nawy, Antonei B. Csoka, Kazuhiro Mio, and Robert Stern Detecting Hyaluronidase and Hyaluronidase Inhibitors: Hyaluronan-Substrate Gel and -Inverse Substrate Gel Techniques Kazuhiro Mio, Antonei B. Csoka, Susan Stair Nawy, and Robert Stern III. Interactions Affinity Coelectrophoresis of Proteoglycan Protein Complexes James D. San Antonio and Arthur D. Lander Binding Constant Measurements for Inhibitors of Growth Factor Binding to Heparan Sulfate Proteoglycans Kimberly E. Forsten and Matthew A. Nugent Interaction of Proteoglycans with Receptor Tyrosine Kinases David K. Moscatello and Renato V. Iozzo Measuring Single-Cell Cytosolic Ca2+ Concentration in Response to Proteoglycans Sandip Patel and Renato V. Iozzo Serum Amyloid A Peptide Interactions with Glycosaminoglycans: Evaluation by Affinity Chromatography John B. Ancsin and Robert Kisilevsky Interactions of Lipoproteins with Proteoglycans Kevin Jon Williams Hyaluronan and Hyaluronan-Binding Proteins: Probes for Specific Detection Bryan P. Toole, Qin Yu, and Charles B. Underhill Novel Confocal-FRAP Analysis of Carbohydrate Protein Interactions Within the Extracellular Matrix Philip Gribbon, Boon C. Heng, and Timothy E. Hardingham Regulatory Roles of Syndecans in Cell Adhesion and Invasion J. Kevin Langford and Ralph D. Sanderson Optical Biosensor Techniques to Analyze Protein Polysaccharide Interactions Dave G. Fernig Phage Display Technology to Obtain Antiheparan Sulfate Antibodies T. H. van Kuppevelt, G. J. Jenniskens, J. H. Veerkamp, G. B. ten Dam, and M. A. B. A. Dennissen Tissue-Specific Binding by FGF and FGF Receptors to Endogenous Heparan Sulfates Andreas Friedl, Mark Filla, and Alan C. Rapraeger

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