Glycobiology Advance Access originally published online on February 17, 2006
Glycobiology 2006 16(6):502-513; doi:10.1093/glycob/cwj093
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A tandem mass spectrometric approach to determination of chondroitin/dermatan sulfate oligosaccharide glycoforms
2 Department of Biochemistry, Boston University School of Medicine, 715 Albany Street, R-806, Boston, MA 02118; and 3 Department of Cell and Molecular Biology, Lund University, BMC B11, S-221 84, Lund, Sweden
1 To whom correspondence should be addressed; e-mail: jzaia{at}bu.edu
Received on October 27, 2005; revised on January 26, 2006; accepted on February 15, 2006
Dermatan sulfate (DS) chains are variants of chondroitin sulfate (CS) that are expressed in mammalian extracellular matrices and are particularly prevalent in skin. DS has been implicated in varied biological processes including wound repair, infection, cardiovascular disease, tumorigenesis, and fibrosis. The biological activities of DS have been attributed to its high content of IdoA(
1–3)GalNAc4S(ß1–4) disaccharide units. Mature CS/DS chains consist of blocks with high and low GlcA/IdoA ratios, and sulfation may occur at the 4- and/or 6-position of GalNAc and 2-position of IdoA. Traditional methods for the analysis of CS/DS chains involve differential digestion with specific chondroitinases followed by steps of chromatographic isolation of the products and di-saccharide analysis on the individual fraction. This work reports the use of tandem mass spectrometry to determine the patterns of sulfation and epimerization of CS/DS oligosaccharides in a single step. The approach is first validated and then applied to a series of skin DS samples and to decorins from three different tissues. DS samples ranged from 74 to 99% of CSB-like repeats, using this approach. Decorin samples ranged from 30% CSB-like repeats for those samples from articular cartilage to 75% for those from sclera. These values agree with known levels of glucuronyl C5-epimerase in these tissues.
Key words: chondroitin sulfate / decorin / dermatan sulfate / glycosaminoglycan / mass spectrometry
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