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.
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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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Zamfir, A.D. et al. (2012). Brain Chondroitin/Dermatan Sulfate, from Cerebral Tissue to Fine Structure: Extraction, Preparation, and Fully Automated Chip-Electrospray Mass Spectrometric Analysis. In: Rédini, F. (eds) Proteoglycans. Methods in Molecular Biology, vol 836. Humana Press. https://doi.org/10.1007/978-1-61779-498-8_10
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DOI: https://doi.org/10.1007/978-1-61779-498-8_10
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