Main description:

This volume is a comprehensive and up-to-date collection of strategies, reproducible methods, and protocols for the in-depth analysis of Proteoglycans (PGs) and their glycan part, the GAGs. Chapters are divided into three parts detailing GAGs in biological specimens, protocols for the evaluation of the in vitro and in vivo effects of PGs/GAGs, and protocols for compounds related with the metabolic enzymes, epigenetic regulation, and PGs/GAGs-based inhibitors. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and methods, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Cutting-edge and thorough, Proteoglycans: Methods and Protocols aims to provide information on the elucidated the structural and functional aspects of the complex matrix macromolecules such as the proteoglycans and glycosaminoglycans.

Contents:

Part I: Glycosaminoglycans: Strategies for Detection and Structure Characterization1. Detection of Glycosaminoglycans in Biological Specimens Shaukat A Kha, FNU Nidhi, Paige C. Amendu, and Shunji Tomatsu
2. Immunochemical Detection and Glycosaminoglycan Disaccharide-Based Characterization Of Chondroitin Sulfate ProteoglycansTadahisa Mikami and Hiroshi Kitagawa
3. Proteoglycans: A Tool for detecting hyaluronan by ELISA-like MethodsDaiana L. Vitale, Rosalia I. Cordo-Russo, and Laura Alaniz
4. Particle Exclusion Assay: A Tool for Measuring Hyaluronan Pericellular MatrixArianna Parnigoni, Paola Moretto, Simona Rovera, Manuela Viola, Evgenia Karousou, Alberto Passi, and Davide Vigetti
5. Urinary Glycosaminoglycans: Characterization and QuantificationNaima Fdil and Es-Said Sabir
6. HPLC-MS characterization of tissue-derived heparan sulfate and chondroitin sulfateGabor Toth, Lilla Turiak
7. Evaluation of Glycosaminoglycans Biological Age in Cells and Tissues Chrysostomi Gialeli and Isabel Goncalves
8. Studying the effects of Glycosaminoglycans in Cell Morphological Aspect with Scanning Electron MicroscopyChristos Koutsakis, Marco Franchi, Anastasia-Gerasimoula Tavianatou, Valentina Masola, and Nikos K. Karamanos
Part II: Proteoglycans: Separation, Analysis, and Interactions9. Evaluation of the In Vitro and In Vivo Effects of Biglycan in Innate ImmunityJinyang Zeng-Brouwers, Lisa Sophie Huber, Rosetta Merline, Jonel Trebicka, Malgorzata Wygrecka, and Liliana Schaefer
10. Analysis of Extracellular Vesicle-associated ProteoglycansJuliana Pocas and Mattias Belting
11. Analysis of Aggrecan Glycanation by Western Blot in Cell CultureChiara Gramegna Tota, Alessandra Leone, Antonio Rossi, and Chiara Paganini
12. Repulsive Scaling Replica Exchange Molecular Dynamics in Modeling Protein-Glycosaminoglycan ComplexesMateusz Marcisz, Martyna Maszota-Zieleniak, and Sergey A. Samsonov
13. In vitro investigation of hyaluronan/CD44 networkTheodore Karalis, Paraskevi Heldin, and Spyridon S. Skandalis
14. Separation and Identification of Native Proteoglycans by Composite Agarose-Polyacrylamide Gel Electrophoresis and ImmunoblottingJames Melrose
15. Building, Visualizing, and Analyzing Glycosaminoglycan-Protein Interaction NetworksSylvie Ricard-Blum
Part III: Proteoglycans related Molecules (Enzymes, Inhibitors, and Epigenetics)16. An Enzymatic Activity Assay for Heparanase That Is Useful for Evaluating Clinically Relevant Inhibitors and Studying KineticsEdward Hammond and Vito Ferro
17. A method for Urinary Trypsin Inhibitor (UTI) purification combining anion-exchange chromatography enrichment and preparative SDS-PAGEGabriele Nieddu, Marilena Formato, and Antonio Junior Lepedda
18. Enzymatic Sequencing of Heparin Oligosaccharides Using ExolyaseQingdong Zhang, Danrong Lu, and Fuchuan Li
19. Evaluating the Effects of microRNAs on Proteoglycans and Matrix Constituents' Expression and Functional PropertiesZoi Piperigkou and Nikos K. Karamanos
20. Proteoglycan Expression Studied by microRNAsNancy Adriana Espinoza-Sanchez, Fabian Troschel, Burkhard Greve, and Martin Goette
21. Modeling and simulations of Extra Cellular GlycoproteinsRajas M Rao, Nicolas Belloy, Jean-Marc Crowet, Manuel Dauchez, and Stephanie Baud