Human follicular fluid heparan sulfate contains abundant 3-O-sulfated chains with anticoagulant activity
Human follicular fluid heparan sulfate contains abundant 3-O-sulfated chains with anticoagulant activity
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DOI：
10.1074/jbc.m805338200
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发表时间：
2008-10-17
影响因子：
4.8
通讯作者：
Linhardt, Robert J.
中科院分区：
文献类型：
--
作者：
de Agostini, Ariane I.;Dong, Ji-Cui;Linhardt, Robert J.
抗凝血剂硫酸乙酰肝素蛋白聚糖通过特异性3-O-硫酸化五糖结合并激活抗凝血酶。它们不仅存在于血管壁中，也存在于血管外组织中，如卵巢，其功能尚不清楚。排卵时卵泡破裂是成年哺乳动物组织重塑的最显著例子之一。它涉及严格控制的炎症，蛋白水解和纤维蛋白沉积。我们假设卵巢硫酸乙酰肝素可能通过与效应蛋白的相互作用来调节这些过程。我们以前的工作表明，抗凝剂硫酸乙酰肝素是由啮齿类动物卵巢颗粒细胞合成的，我们现在已经开始从人类卵泡液中表征硫酸乙酰肝素。在这里，我们报告了第一个抗凝剂硫酸乙酰肝素纯化的天然人血管外来源。硫酸乙酰肝素链根据其对抗凝血酶的亲和力进行分级分离，并通过H-1 NMR和MS/MS分析其结构。我们发现，人卵泡液是一个丰富的来源，抗凝剂硫酸乙酰肝素，占总硫酸乙酰肝素的50.4%。这些抗凝血酶结合链含有超过6%的3-O-硫酸葡糖胺残基，对人卵泡液的抗凝活性为2.5 IU/ml，抗Xa因子比活性为167 IU/mg。不结合抗凝血酶的硫酸乙酰肝素链令人惊讶地显示出极高的3-O-硫酸化葡糖胺残基含量，这表明它们可能通过与其他蛋白质的相互作用而显示出生物活性。
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 H-1 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.