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Proteoglycans pp 145-159 | Cite as

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

  • Alina D. ZamfirEmail author
  • Corina Flangea
  • Alina Serb
  • Eugen Sisu
  • Leon Zagrean
  • Andreas Rizzi
  • Daniela G. Seidler
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 836)

Abstract

Chondroitin sulfate (CS) and dermatan sulfate (DS) glycosaminoglycans (GAGs) are covalently linked to proteins, building up a wide range of proteoglycans, with a prevalent expression in the extracellular matrix (ECM). In mammalian tissues, these GAG species are often found as hybrid CS/DS chains. Their structural diversity during chain elongation is produced by variability of sulfation in the repeating disaccharide units. In central nervous system, a large proportion of the ECM is composed of proteoglycans; therefore, CS/DS play a significant role in the functional diversity of neurons, brain development, and some brain diseases. A requirement for collecting consistent data on brain proteoglycan glycosylation is the development of adequate protocols for CS/DS extraction and detailed compositional and structure analysis. This chapter will present a strategy, which combines biochemical tools for brain CS/DS extraction, purification, and fractionation, with a modern analytical platform based on chip-nanoelectrospray multistage mass spectrometry (MS) able to provide information on the essential structural elements such as epimerization, chain length, sulfate content, and sulfation sites.

Key words

Glycosaminoglycan Chondroitin sulfate Dermatan sulfate Cerebral tissue Chip-nanoelectrospray Multistage mass spectrometry 

Notes

Acknowledgments

The authors are grateful to the Romanian National Authority for Scientific Research (Grants No. PN-II-RU-TE-2011-2-0008 and PN- II-ID-PCE-2011-3-0047) and German Society for Research (DFG SE1431/1-1 and IRTG1549/1 Molecular and Cellular Glycosciences) for financial support.

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Alina D. Zamfir
    • 1
    • 2
    Email author
  • Corina Flangea
    • 1
  • Alina Serb
    • 3
  • Eugen Sisu
    • 3
  • Leon Zagrean
    • 4
  • Andreas Rizzi
    • 5
  • Daniela G. Seidler
    • 6
  1. 1.Department of Chemical and Biological Sciences“Aurel Vlaicu” University of AradAradRomania
  2. 2.Mass Spectrometry LaboratoryNational Institute for Research and Development in Electrochemistry and Condensed MatterTimisoaraRomania
  3. 3.Department of Biochemistry“Victor Babes” University of Medicine and PharmacyTimisoaraRomania
  4. 4.Neuroscience Laboratory“Carol Davila” University of Medicine and PharmacyBucharestRomania
  5. 5.Institute of Analytical Chemistry and Food ChemistryUniversity of ViennaViennaAustria
  6. 6.Institute for Physiological Chemistry and PathobiochemistryUniversity Hospital of MünsterMünsterGermany

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