Proteoglycan Protocols

ISBN-13: 9781617371745 / Angielski / Miękka / 2010 / 558 str.

Renato V. Iozzo

Opis

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.

Recenzje

"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

Spis treści

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 SvenBjörnsson 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

Blurb

Proteoglycans, highly complex macromolecules found in mammalian and lower organisms, serve a multitude of biological functions. In Proteoglycan Protocols, Renato Iozzo, MD, 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. Each method is carefully described by leading experts in the field and includes practical considerations that are often lacking in formal papers.

Authoritative and detailed, Proteoglycan Protocols offers molecular and cellular biologists, biochemists, pathologists, and geneticists a comprehensive reference and practical guide to the most common methods used today for the isolation and characterization of proteoglycans from cultured cells, tissues, and biological fluids.