Mapping and quantification of the major oligosaccharide components of heparin

Abstract

A new method of determining the oligosaccharide composition of commercial glycosaminoglycan heparin is described in which heparin was first depolymerized using heparin lyase (EC 4.2.2.7), and then analysed by a single h.p.l.c. step. All 20 of the porcine and bovine heparins examined were found to contain a small number of major oligosaccharide components, which on average comprised 86% of their mass. The five most abundant oligosaccharides have defined chemical structures. Although the relative abundance of oligosaccharides varied, the heparins examined were surprisingly similar. Porcine, bovine, low-Mr, and high and low antithrombin III (ATIII)-affinity heparins, however, each had distinctly different proportions of these major oligosaccharide components. The concentrations of one of these five oligosaccharides, containing a portion of the ATIII binding site, correlated with the anticoagulant activity of the ATIII-affinity-fractionated porcine-mucosal heparins from which it was derived. An additional oligosaccharide of undetermined structure was found in significant quantities in both bovine heparin and high ATIII-affinity porcine-mucosal heparin. The correlation between oligosaccharide concentration and anticoagulant activity suggests that the oligosaccharide is derived from a structural variant of the ATIII-binding site. Finally, for the heparins examined chondroitin/dermatan sulphate formed 0.6-7.4% of their mass.

The Retinoic Acid and cAMP-dependent Up-regulation of 3-O-Sulfotransferase-1 Leads to a Dramatic Augmentation of Anticoagulantly Active Heparan Sulfate Biosynthesis in F9 Embryonal Carcinoma Cells

“…Thus, the RACT-induced F9 cell is similar to the mast cell, which contains a large proportion of molecules with the AT binding property (30%) (2). Structural and biochemical analysis also indicates that the precursor of anticoagulant heparin in mast cells might be present in only small amounts (28,29).…”

Effects of Structural Modification of Calcium Spirulan, a Sulfated Polysaccharide from Spirulina Platensis, on Antiviral Activity.

“…6) To obtain depolymerized derivatives of the sulfated polysaccharides heparin and fucoidan, chemical methods [7][8][9] and enzymatic reaction systems 10,11) have been used to cleave the glycosidic bonds of the polysaccharides.…”

Expression of Heparan Sulfate d-Glucosaminyl 3-O-Sulfotransferase Isoforms Reveals Novel Substrate Specificities

“…Indeed, the available data support this hypothesis (16 -18). As documented by several researchers including ourselves, 3-O-sulfation of glucosamine is a rare modification that constitutes the last step in the biosynthesis of HS (14,15,19,20). We have shown that 3-OST-1 requires an appropriate HS precursor with defined monosaccharide sequence for sulfation to occur and make HS act .…”