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Glycobiology Advance Access published online on December 22, 2007
Glycobiology, 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

Anti-Thrombin Activity and Disaccharide Composition of Dermatan Sulphate from Different Bovine Tissues

Simone A. Osborne1,2,*, Robyn A. Daniel1,2,, Kirthi Desilva2,3, and Robert Seymour1,2,

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

* To whom correspondence should be addressed. Phone 617-3214 2274, Fax 617-3214 2900, Simone.Osborne{at}csiro.au

Received on September 21, 2007; accepted on December 13, 2007

Dermatan sulphate is a glycosaminoglycan that selectively inhibits the action of thrombin through 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 sulphate an attractive candidate as an anti-thrombotic drug. Previous studies have showed that dermatan sulphate 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 sulphate derived from mammalian tissues other than intestinal mucosa or skin. Ten different bovine tissues including the aorta, diaphragm, eyes, large and small intestine, oesophagus, skin, tendon, tongue and tongue skin were used to prepare dermatan sulphate enriched fractions by anion exchange chromatography and acetone precipitation. Heparin cofactor II/dermatan sulphate mediated thrombin inhibition measured in vitro revealed activity comparable to or higher than the commercial standard with two-fold differences observed between some tissues. Analysis of the extracted dermatan sulphate using fluorophore-assisted carbohydrate electrophoresis revealed significant differences in the relative percentage of all the mono-sulphated disaccharides, in particular the predominant mammalian disaccharide uronic acid->N-acetyl-D-galactosamine-4-O-sulphate, confirming previous reports regarding variations in sulphation in dermatan sulphate from different tissues. Overall, these findings demonstrate that dermatan sulphate extracted from a range of bovine tissue exhibits in vitro anti-thrombin activity equivalent to or higher than that observed for porcine intestinal mucosa, identifying additional sources of dermatan sulphate as potential anti-thrombotic agents.

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


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