Glycobiology Advance Access originally published online on June 10, 2003
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Glycobiology, 2003, Vol. 13, No. 11 733-742
© 2003 Oxford University Press
Structural investigation of chondroitin/dermatan sulfate oligosaccharides from human skin fibroblast decorin
2,33 Institute for Medical Physics and Biophysics, Biomedical Analysis Department, University of Münster, Robert-Koch-Str. 31, D-48149, Münster, Germany; 4 National Institute for Research and Development in Electrochemistry and Condensed Matter, Timisoara, Romania; and 5 Institute of Physiological Chemistry and Pathobiochemistry, University of Münster, Waldeyerstr. 15, D-48149, Münster, Germany
Received on November 15, 2002; revised on May 22, 2003; accepted on May 22, 2003
Hybrid chondroitin/dermatan sulfate (CS/DS) glycosaminoglycan chains, derived from decorin secreted by human skin fibroblasts, were shown to interact with FGF-2, as did oligosaccharides derived therefrom by chondroitin B lyase digestion. In a first attempt to identify the biologically active sequence, a novel protocol for structural analysis of enzyme-resistant oligosaccharides larger than standard trisulfated hexasaccharides was developed. The method bases on capillary electrophoresis (CE) for separating oversulfated species in offline combination with nanoelectrospray ionization quadrupole time-of-flight tandem mass spectrometry (nanoESI-QTOF-MS/MS) in the negative ion mode. Under optimized CE and ESI-MS conditions, up to 12-mer oligosaccharides with different degrees of sulfation were identified. A novel tandem MS protocol (CID-VE) was applied to elucidate the structure of a previously undescribed pentasulfated CS/DS hexasaccharide, 
-4,5-IdoAGalNAc[GlcAGalNAc]2(5S). In this molecular species, detected as a triply charged ion at m/z 511.38, three sulfates are found in the IdoAGalNAcGlcA moiety offering two structural variants: one containing sulfated IdoA together with a disulfated GalNAc moiety and in the other one both uronic acids, that is, GlcA and IdoA and the amino sugar each carry a sulfate ester group.
1 These authors contributed equally to this work
2 To whom the correspondence should be addressed; email: jkp{at}uni-muenster.de
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