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Glycobiology Advance Access published online on February 17, 2006
Glycobiology, doi:10.1093/glycob/cwj093
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© The Author 2006. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org
Received October 27, 2005
Revised January 26, 2006
Accepted February 15, 2006

Article

A tandem mass spectrometric approach to determination of chondroitin/dermatan sulfate oligosaccharide glycoforms

May Joy C. Miller 1, Catherine E. Costello 1, Anders Malmström 2, and Joseph Zaia 1 *

1 Department of Biochemistry, Boston University School of Medicine, Boston, MA
2 Department of Cell and Molecular Biology, Lund University, Lund, Sweden

* To whom correspondence should be addressed.
Joseph Zaia, E-mail: jzaia{at}bu.edu


  Abstract

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({alpha}1-3)GalNAc4S({beta}1-4) 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 analysis of CS/DS chains involve differential digestion with specific chondroitinases followed by steps of chromatographic isolation of the products and disaccharide analysis on the individual fraction. This work reports the use of tandem mass spectrometry to determine 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 74-99% of CSB-like repeats, using this approach. Decorin samples ranged from 30% CSB-like repeats for that from articular cartilage to 75% for that from sclera. These values agree with known levels of glucuronyl C5-epimerase in these tissues.

Keywords: chondroitin sulfate/decorin/dermatan sulfate/glycosaminoglycan/mass spectrometry.
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