Open AccessSynthesis of 3-O-Sulfated Disaccharide and Tetrasaccharide Standards for Compositional Analysis of Heparan Sulfate
Author(s) -
Vijay M. Dhurandhare,
Vijayakanth Pagadala,
Andreia Ferreira,
Louis De Muynck,
Jian Liu
Publication year - 2019
Publication title -
biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.43
H-Index - 253
eISSN - 1520-4995
pISSN - 0006-2960
DOI - 10.1021/acs.biochem.9b00838
Subject(s) - tetrasaccharide , sulfation , disaccharide , chemistry , oligosaccharide , iduronic acid , depolymerization , glucosamine , heparan sulfate , biochemistry , uronic acid , glycosaminoglycan , sulfate , residue (chemistry) , stereochemistry , organic chemistry , polysaccharide
3- O -Sulfation on the glucosamine sugar unit in heparan sulfate (HS) is linked to various biological functions, including the anticoagulant activity to treat thrombotic disorders in hospitals. The 3- O -sulfated glucosamine is biosynthesized by heparan sulfate glucosamine 3-sulfotransferases. Because of its biological significance, there is a need for 3- O -sulfated oligosaccharide standards to facilitate the compositional analysis of HS. These oligosaccharides must contain a Δ 4,5 -unsaturated uronic acid (ΔUA) residue at the nonreducing end, which is due to the depolymerization reaction catalyzed by heparin lyases used during the compositional analysis procedure. Here, we describe a protocol for the preparation of one 3- O -sulfated disaccharide (compound 4 ) and three 3- O -sulfated tetrasaccharides (compound 1 - 3 ) in a milligram scale. The synthesis of 3- O -sulfated disaccharide and tetrasaccharide standards was completed by degrading synthetic octasaccharides using heparin lyases. Further analysis revealed that 3- O -sulfated oligosaccharide standards are labile under basic conditions, confirming the findings from a previous study. The unwanted degradation was reduced by decreasing the pH in the presence of phosphate buffer. The 3- O -sulfated oligosaccharide standards are reagents to characterize 3- O -sulfation in HS derived from biological sources.