Antithrombin activity and disaccharide composition of dermatan sulfate from different bovine tissues

Abstract

Dermatan sulfate is a glycosaminoglycan that selectively inhibits the action of thrombin through interaction with heparin cofactor II. Unlike heparin it does not interact with other coagulation factors and is able to inhibit thrombin associated with clots. This property has made dermatan sulfate an attractive candidate as an antithrombotic drug. Previous studies have showed that dermatan sulfate derived from porcine/bovine intestinal mucosa/skin or marine invertebrates is capable of stimulating heparin cofactor II-mediated thrombin inhibition in vitro. This biological activity is reported for the first time in this study using dermatan sulfate derived from mammalian tissues other than intestinal mucosa or skin. Ten different bovine tissues including the aorta, diaphragm, eyes, large and small intestine, esophagus, skin, tendon, tongue, and tongue skin were used to prepare dermatan sulfate-enriched fractions by anion exchange chromatography and acetone precipitation. Heparin cofactor II/dermatan sulfate-mediated thrombin inhibition measured in vitro revealed activity comparable to or higher than the commercial standard with 2-fold differences observed between some tissues. Analysis of the extracted dermatan sulfate using fluorophore-assisted carbohydrate electrophoresis revealed significant differences in the relative percentage of all the mono-sulfated disaccharides, in particular the predominant mammalian disaccharide uronic acid-->N-acetyl-D-galactosamine-4-O-sulfate, confirming previous reports regarding variations in sulfation in dermatan sulfate from different tissues. Overall, these findings demonstrate that dermatan sulfate extracted from a range of bovine tissues exhibits in vitro antithrombin activity equivalent to or higher than that observed for porcine intestinal mucosa, identifying additional sources of dermatan sulfate as potential antithrombotic agents.

Grafting of dermatan sulfate on polyethylene terephtalate to enhance biointegration

“…DS is an ubiquitous element of the extracellular matrix. This glycosaminoglycan is composed of a linear polysaccharide chain assembled from disaccharide units containing a hexosamine, N‐acetyl galactosamine (GalNAc) and a hexuronic acid, iduronic (Ido A), or glucuronic acid (Glc A) 20. We previously isolated a DS from the skin of the shark, Scyliorhinus canicula , with high anticoagulant activity due to the enhancement of thrombin inhibition by heparin cofactor II (HCII) but without effect on platelet aggregation and activation 21.…”

Automated Chemical Oligosaccharide Synthesis: Novel Approach to Traditional Challenges

“…The second target was dermatan sulfate, which is a polysaccharide composed by a disaccharide repeating unit, consisting of N -acetyl- β - d -galactosamine and l -iduronic acid. 565 Seeberger et al developed a synthesis of a di- and a tetrasaccharide using the Fmoc protection at the oligosaccharide propagation position and the Lev ester at the future sulfation sites. Galactosamine is readily available in a small number of steps, while iduronic acid requires a more complex synthesis.…”

The Sea as a Rich Source of Structurally Unique Glycosaminoglycans and Mimetics

“…Although the sulfation pattern of dermatan sulfate, which is mostly composed of disaccharide building block [→4)-IdoA-(α1→3)-GalNAc4S-(β1→], can vary among tissues [ 44 ], cells [ 45 ] and pathophysiological conditions [ 46 ]; the composing GalNAc units are mostly sulfated at the C4 position (~95%) as shown in Figure 1 D. But minor amounts of sulfation (~15%) can also happen at different positions such as the C6 position of GalNAc and (~10%) at the C2 position of the composing IdoA. In ascidians (Urochordata, Ascidiaceae), also known as tunicates or sea squirts, different sulfation patterns can be observed on their dermatan sulfates ( Figure 2 C,D) [ 42 , 43 , 47 ].…”