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Glycobiology Advance Access originally published online on December 22, 2007
Glycobiology 2008 18(3):225-234; doi:10.1093/glycob/cwm136
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© The Author 2007. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: journals.permissions@oxfordjournals.org

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

Simone A. Osborne1,2,3,, Robyn A. Daniel2,3,, Kirthi Desilva3,4, and Robert B. Seymour2,3,

2 CSIRO, Livestock Industries, Queensland Bioscience Precinct, 306 Carmody Road, St Lucia, Queensland 4067
3 CSIRO, Food Futures Flagship, 5 Julius Ave, Riverside Corporate Park, North Ryde, New South Wales 2113
4 CSIRO, Food Science Australia, 671 Sneydes Road, Werribee, Victoria 3030, Australia

1 To whom correspondence should be addressed: Tel: +617-3214-2274; Fax: +617-3214-2900; e-mail: Simone.Osborne{at}csiro.au

Received on September 21, 2007; revised on November 16, 2007; accepted on December 13, 2007

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.

Key words: Bovine / dermatan sulfate / disaccharide / heparin cofactor II / thrombin inhibition


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