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Main description:
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.
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 SvenBjoernsson
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
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: November, 2010
Pages: 576
Weight: 924g
Availability: Available
Subcategories: Biochemistry
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