The 3--sulfated glucosamine in heparan sulfate (HS) is a low-abundance structural component, but it is a key saccharide unit for the biological activities of HS. A method to determine the level of 3--sulfated HS is lacking. Here, we describe a LC-MS/MS based method to analyze the structural motifs. We determined the levels of 3--sulfated structural motifs from pharmaceutical heparin manufactured from bovine, porcine, and ovine. We discovered that saccharide chains carrying 3--sulfation from enoxaparin, an FDA-approved low-molecular weight heparin, displayed a slower clearance rate than non-3--sulfated sugar chains in a mouse model. Lastly, we detected the 3--sulfated HS from human brain. Furthermore, we found that a specific 3--sulfated structural motif, tetra-1, is elevated in the brain HS from Alzheimer's disease patients ( = 5, = 0.0020). Our method offers a practical solution to measure 3--sulfated HS from biological sources with the sensitivity and quantitative capability.
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| http://dx.doi.org/10.1021/acs.analchem.1c04965 | DOI Listing |
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