Main description:

Now recognized as a reservoir for growth factors and cytokines modulating cell activation status and turnover, proteoglycans have stimulated great amount of interest and research. In Proteoglycans: Methods and Protocols, experts in the field examine issues of basic concepts and up-to-date analysis methods for proteoglycan and glycosaminoglycan (GAG) at the protein and saccharide levels. The volume continues with the multifunctional aspect of proteoglycans, highlighted through three relevant examples of proteoglycans with highly different strucutures: serglycin, aggrecan, and heparin sulphate proteoglycans. The final chapter describes proteoglycan involvement in the pathogenesis of various disorders and their potential therapeutic value in osteo-articular diseases. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and accessible, Proteoglycans: Methods and Protocols is an ideal guide for scientists attempting to pursue further research in this vital and complex area of study.

Contents:

Part I: Proteoglycans

1. Proteoglycans: Gene Cloning

Mauricio Cortes, James R. Mensch, Miriam Domowicz, and Nancy B. Schwartz

2. Proteoglycan: Site Mapping and Site-Directed Mutagenesis

Fred K. Hagen

3. Mapping of the Wnt/ -Catenin/TCF Response Elements in the Human Versican Promoter

Maziar Rahmani, Jon M. Carthy, and Bruce M. McManus

4. Gene Silencing in Mouse Embryonic Stem Cells

Norihiko Sasaki and Shoko Nishihara

5. A Novel Strategy for a Splice-Variant Selective Gene Ablation: The Example of the Versican V0/V2 Knockout

Maria T. Dours-Zimmermann and Dieter R. Zimmermann

6. Detection of Neurocan in Cerebrospinal Fluid

Uwe Rauch

Part II: Glycosaminoglycans Analysis

7. Glycosaminoglycan Chain Analysis and Characterization (Glycosylation/Epimerization)

Shuji Mizumoto and Kazuyuki Sugahara

8. Characterization of Glycosaminoglycans by Tandem Vibrational Microspectroscopy and Multivariate Data Analysis

Nathalie Mainreck, Stephane Brezillon, Ganesh D. Sockalingum, Francois-Xavier Maquart, Michel Manfait, and Yanusz Wegrowski

9. Glycosaminoglycans: Oligosaccharide Analysis by Liquid Chromatography/Specific Labeling

Derek J. Langelsay, Christopher J. Jones, Szabolcs Beni, and Cynthia K. Larive

10. Brain Chondroitin/Dermatan Sulfate, from Cerebral Tissue to Fine Structure: Extraction, Preparation, and Fully Automated Chip-Electrospray Mass Spectrometric Analysis

Alina D. Zamfir, Corina Flangea, Alina Serb, Eugen Sisu, Leon Zagrean, Andreas Rizzi, and Daniela G. Seidler

11. Use of Neutrons Reveal the Dynamics of Cell Surface Glycosaminoglycans

Marion Jasnin

12. Following Protein-Glycosaminoglycan (GAG) Polysaccharide Interactions with Differential Scanning Fluorimetry (DSF)

Katarzyna A. Uniewicz, Alessandro Ori, Timothy R. Rudd, Marco Guerrini, Mark C. Wilkinson, David G. Fernig, and Edwin A. Yates

13. In Vivo Scintigraphic Imaging of Proteoglycans

Elisabeth Miot-Noirault, Aurelien Vidal, Philippe Auzeloux, Caroline Peyrode, Jean-Claude Madelmont, and Jean-Michel Chezal

Part III: PGs: Multifunctional Cell Regulators

14. Serglycin: The Master of the Mast Cell

Elin Roennberg and Gunnar Pejler

15. Analysis of Aggrecan Catabolism by Immunoblotting and Immunohistochemistry

Peter J. Roughley and John S. Mort

16. Heparan Sulfate Proteoglycans as Multifunctional Cell Regulators: Cell Surface Receptors

Jin-ping Li and Dorothe Spillmann

Part IV: Proteoglycans Involvement in Pathophysiology

17. Models for Studies of Proteoglycans in Kidney Pathophysiology

Scott J. Harvey

18. Lumican Promotes Corneal Epithelial Wound Healing

Chia-Yang Liu and Winston Whei-Yang Kao

19. Shedding of Cell Membrane-Bound Proteoglycans

Eon Jeong Nam and Pyong Woo Park

20. Modulatory Effects of Proteoglycans on Proteinase Activities

Steven Georges, Dominique Heymann, and Marc Padrines

21. Proteoglycans and Osteolysis

Marc Baud'Huin, Celine Charrier, Gwenola Bougras, Regis Brion, Frederic Lezot, Marc Padrines, and Dominique Heymann

22. Proteoglycans and Cartilage Repair

Mohamed Ouzzine, Narayanan Venkatesan, and Sylvie Fournel-Gigleux