Unfolded von Willebrand factor binds protein S and reduces anticoagulant activity

Blood Vessels, Thrombosis & Hemostasis Pub Date : 2025-02-01 DOI:10.1016/j.bvth.2024.100030

Martha M. S. Sim , Molly Y. Mollica , Hammodah R. Alfar , Melissa Hollifield , Dominic W. Chung , Xiaoyun Fu , Siva Gandhapudi , Daniëlle M. Coenen , Kanakanagavalli Shravani Prakhya , Dlovan F. D Mahmood , Meenakshi Banerjee , Chi Peng , Xian Li , Alice C. Thornton , James Z. Porterfield , Jamie L. Sturgill , Gail A. Sievert , Marietta Barton-Baxter , Ze Zheng , Kenneth S. Campbell , Jeremy P. Wood

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Abstract

The critical plasma anticoagulant protein S (PS) circulates in 2 functionally distinct pools: free (anticoagulant) or bound to complement component 4b-binding protein (C4BP; anti-inflammatory). Acquired free PS deficiency is detected in several viral infections, but its cause is unclear. Here, we used biochemical approaches and human patient plasma samples to identify an interaction between PS and von Willebrand factor (VWF), which causes free PS deficiency and reduced PS anticoagulant activity. We first identified a shear-dependent interaction between PS and VWF by mass spectrometry. Consistently, PS and VWF could be crosslinked together in plasma, and plasma PS and VWF comigrated in gel electrophoresis. The PS/VWF interaction was blocked by and tissue factor pathway inhibitor but not activated protein C, suggesting an interaction with the sex hormone binding globulin region of PS. Microfluidic systems demonstrated that PS stably binds VWF as VWF unfolds under turbulent flow. PS/VWF complexes also localized to platelet thrombi under laminar arterial flow. In thrombin generation–based assays, shearing plasma decreased PS activity, an effect not seen in the absence of VWF. Finally, free PS deficiency in patients with COVID-19 correlated with changes in VWF, but not C4BP, and with thrombin generation. Our data indicate that PS binds to a shear-exposed site on VWF, thus sequestering free PS and decreasing its anticoagulant activity, which would account for the increased thrombin generation potential. Because many viral infections present with free PS deficiency, elevated circulating VWF, and increased vascular shear, we propose that the PS/VWF interaction reported here is a likely contributor to virus-associated thrombotic risk.

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未折叠血管性血友病因子结合蛋白S降低抗凝血活性

关键血浆抗凝蛋白S （PS）在两个功能不同的池中循环：游离（抗凝）或与补体组分4b结合蛋白（C4BP）结合；抗炎)。在几种病毒感染中发现获得性游离PS缺陷，但其原因尚不清楚。在这里，我们使用生化方法和人类患者血浆样本来确定PS和血管性血友病因子（VWF）之间的相互作用，导致游离PS缺乏和降低PS抗凝血活性。我们首先通过质谱法确定了PS和VWF之间的剪切依赖相互作用。PS和VWF在血浆中可以交联在一起，血浆PS和VWF在凝胶电泳中可以同质。PS/VWF的相互作用被组织因子途径抑制剂阻断，但蛋白C未被激活，提示与PS的性激素结合球蛋白区相互作用。微流控系统显示，当VWF在湍流中展开时，PS稳定地与VWF结合。PS/VWF复合物也定位于层动脉血流下的血小板血栓。在以凝血酶生成为基础的实验中，剪切血浆降低了PS活性，这种效应在没有VWF的情况下没有观察到。最后，COVID-19患者游离PS缺乏与VWF变化相关，但与C4BP和凝血酶生成无关。我们的数据表明，PS与VWF上剪切暴露的位点结合，从而隔离游离PS并降低其抗凝血活性，这可能是导致凝血酶生成潜力增加的原因。由于许多病毒感染表现为游离PS缺乏、循环VWF升高和血管剪切增加，我们认为这里报道的PS/VWF相互作用可能是病毒相关血栓形成风险的一个因素。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Blood Vessels, Thrombosis & Hemostasis

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