Human follicular fluid heparan sulfate contains abundant 3-O-sulfated chains with anticoagulant activity.

Document Type

Journal Article

Publication Date

10-17-2008

Journal

The Journal of biological chemistry

Volume

283

Issue

42

DOI

10.1074/jbc.M805338200

Keywords

Anticoagulants; Chromatography, Gel; Chromatography, Ion Exchange; Female; Follicular Fluid; Heparitin Sulfate; Humans; Inflammation; Magnetic Resonance Spectroscopy; Mass Spectrometry; Models, Biological; Models, Chemical; Ovary; Sulfur; Sulfuric Acid Esters

Abstract

Anticoagulant heparan sulfate proteoglycans bind and activate antithrombin by virtue of a specific 3-O-sulfated pentasaccharide. They not only occur in the vascular wall but also in extravascular tissues, such as the ovary, where their functions remain unknown. The rupture of the ovarian follicle at ovulation is one of the most striking examples of tissue remodeling in adult mammals. It involves tightly controlled inflammation, proteolysis, and fibrin deposition. We hypothesized that ovarian heparan sulfates may modulate these processes through interactions with effector proteins. Our previous work has shown that anticoagulant heparan sulfates are synthesized by rodent ovarian granulosa cells, and we now have set out to characterize heparan sulfates from human follicular fluid. Here we report the first anticoagulant heparan sulfate purified from a natural human extravascular source. Heparan sulfate chains were fractionated according to their affinity for antithrombin, and their structure was analyzed by 1H NMR and MS/MS. We find that human follicular fluid is a rich source of anticoagulant heparan sulfate, comprising 50.4% of total heparan sulfate. These antithrombin-binding chains contain more than 6% 3-O-sulfated glucosamine residues, convey an anticoagulant activity of 2.5 IU/ml to human follicular fluid, and have an anti-Factor Xa specific activity of 167 IU/mg. The heparan sulfate chains that do not bind antithrombin surprisingly exhibit an extremely high content in 3-O-sulfated glucosamine residues, which suggest that they may exhibit biological activities through interactions with other proteins.

Comments

This is an open access PubMed Central article.

Peer Reviewed

1

Open Access

1

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