von Willebrand Factor without the A2 Domain Is Resistant to Proteolysis

Lankhof, H.; Damas, C; Schiphorst, ME; Mj, IJsseldijk; Bracke, Madelon; Furlan, Miha; Hm, Tsai; Groot, de; Sixma, JJ; Vink, Tom

doi:10.1055/s-0038-1656094

Cited by 30 publications

(32 citation statements)

References 28 publications

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“…wt-VWF, VWF/R1205H, and VWF/D2509G were purified from conditioned serum-free medium (Ultroser G from Biosepra, Cergy-Saint Christophe, France) by immunoaffinity chromatography employing the antibody RU-8 as described (30). The truncated variants VWF/A1-A3, VWF/DЈ-D3, VWF/DЈ-A3, and VWF/A1-CK were purified from conditioned serum-free medium by nickel-nitrilotriacetic acid chromatography as instructed by the manufacturer (Qiagen).…”

Section: Methodsmentioning

confidence: 99%

An Experimental Model to Study the in Vivo Survival of von Willebrand Factor

Lenting

Westein

Terraube

et al. 2004

Journal of Biological Chemistry

140

189

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To explore the molecular basis of von Willebrand factor (VWF) clearance, an experimental model employing VWF-deficient mice was developed. Biodistribution was examined by the injection of radiolabeled VWF, which was primarily directed to the liver with minor amounts in other organs. Disappearance of VWF from plasma was characterized by a rapid initial phase (t1 ⁄2 ␣ ‫؍‬ 13 min) and a slow secondary phase (t1 ⁄2 ␤ ‫؍‬ 3 h), with a mean residence time (MRT) of 2.8 h. A similar clearance was observed for VWF consisting of only high or low molecular weight multimers, indicating that, in our experimental model, clearance is independent of multimeric distribution. This allowed us to compare the survival of fulllength VWF to truncated variants. Deletion of both the amino-terminal D -D3 and carboxyl-terminal D4-CK domains resulted in a fragment with a similar clearance to wild-type VWF. Deletion of only the D -D3 region was associated with an almost 2-fold lower recovery and increased clearance (MRT ‫؍‬ 1.6 h), whereas deletion of only the D4-CK region resulted in a significantly reduced clearance (MRT ‫؍‬ 4.5 h, p < 0.02). These results point to a role of the D -D3 region in preventing clearance of VWF. Furthermore, replacement of D3 domain residue Arg-1205 by His resulted in a markedly increased clearance (MRT ‫؍‬ 0.3 h; p ‫؍‬ 0.004). Therefore, this mutation seems to abrogate the protective effect of the D -D3 region. In vitro analysis of this mutant also revealed a 2-fold reduced affinity for VWF propeptide at low pH, showing that mutation of Arg-1205 results not only in an increased clearance rate but is also associated with an impaired pH-dependent interaction with VWF propeptide.von Willebrand factor (VWF) 1 is a multimeric plasma protein that participates in the hemostatic process. The absence of functional VWF is associated with an abnormal bleeding tendency known as von Willebrand Disease (VWD) (1, 2). VWF contributes to hemostasis in a dual manner. First, it promotes the adhesion of platelets at sites of vascular injury by acting as a molecular bridge between the sub-endothelial collagen matrix and the platelet-surface receptor complex glycoprotein (Gp) Ib␣/IX/V (3). In addition, VWF serves as a carrier protein for coagulation factor VIII (FVIII) in the circulation. This chaperone function results in stabilization of the FVIII heterodimeric structure (4) and protection of FVIII from premature clearance by the low density lipoprotein receptor-related protein (5, 6).VWF is produced and stored in endothelial cells and megakaryocytes. It is synthesized as pre-pro-VWF, a single chain polypeptide with the domain structure

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Section: Methodsmentioning

confidence: 99%

An Experimental Model to Study the in Vivo Survival of von Willebrand Factor

Lenting

Westein

Terraube

et al. 2004

Journal of Biological Chemistry

140

189

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“…[19][20][21] Plasma-derived (pd)-VWF was a kind gift from C. Mazurier (LFB, Lille, France). The ␣M I-domain fused to glutathione-S-transferase (GST) was expressed and purified as follows: cDNA encoding residues Ser112-Gly321 of the ␣M subunit was obtained by standard molecular biology procedures using U937-derived mRNA as source material.…”

Section: Proteinsmentioning

confidence: 99%

P-selectin glycoprotein ligand 1 and β2-integrins cooperate in the adhesion of leukocytes to von Willebrand factor

Pendu

Terraube

Olivier

et al. 2006

Blood

162

159

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Von Willebrand factor (VWF) is an essential component of hemostasis. However, animal studies using VWF-deficient mice suggest that VWF may also contribute to inflammation. In the present study, we demonstrate that VWF was able to interact with polymorphonuclear leukocytes (PMNs) and monocytes under static and flow conditions. Adhesion under flow was dominated by short-lasting contact with resting PMNs, whereas adhesion of phorbol-12-myristate-13-acetate (PMA)-stimulated PMNs was characterized by firm adhesion. Transient binding of PMNs to VWF appeared to be mediated by P-selectin glycoprotein ligand-1 (PSGL-1). Moreover, recombinant PSGL-1 protein and cell surface-expressed PSGL-1 directly interacted with VWF. As for stable adhesion by PMA-stimulated PMNs, we observed that static adhesion and adhesion under flow were strongly inhibited (greater than 75%) by neutrophilinhibitory factor, an inhibitor of ␤2-integrin function. In addition, the isolated I-domain of ␣M␤2 bound to VWF, and cell lines expressing ␣L␤2 or ␣X␤2 adhered efficiently to VWF. Taken together, our data showed that VWF can function as an adhesive surface for various leukocyte subsets (monocytes, PMNs). Analogous to VWF-platelet interaction, VWF provided binding sites for leukocyte receptors involved in rolling (PSGL-1) and stable (␤2-integrins) adhesion. VWF is unique in its intrinsic capacity to combine the rolling and the stable adhesion step in the interaction with leukocytes. IntroductionRecruiting leukocytes to sites of infection or tissue damage is a key element in the inflammatory response. However, these inflammatory cells are in a resting, low-adhesive state while circulating in blood or lymphatic vessels. Stimulatory signals are therefore required to promote the migration of leukocytes through the endothelial layer. This migration involves a complex process that distinguishes a number of distinct steps, including initial rolling to slow down the leukocytes and firm adhesion to allow transendothelial migration. 1 Several endothelial and leukocyte receptors have been identified that contribute to these processes. For instance, leukocyte rolling over the endothelial layer is predominantly mediated through interactions between selectins that are exposed on the stimulated endothelial cell and the P-selectin glycoprotein ligand-1 (PSGL-1) on the leukocyte surface. 2 These interactions result not only in the deceleration of cells but in the activation of signaling pathways that bring these leukocytes into the highadhesive state required for stable adhesion. 3 Important players in the stable adhesion step are the ␤2-integrins. This leukocytespecific integrin family consists of 4 isotypes (␣L␤2, ␣M␤2, ␣D␤2, and ␣X␤2), each of which has the intrinsic capacity to mediate stable adhesion. 4 The major counterreceptors for the ␤2-integrin isotypes are the intercellular adhesion molecules (ICAMs). 4 It has become clear, however, that many other proteins have the capacity to interact with this integrin family, including fibrinogen, vitronectin, juncti...

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“…This interaction is responsible for the tethering, rolling, and activation of platelets that eventually become firmly adhered, leading to thrombus formation within a coronary artery. 3,4 Mature VWF consists of a 2050-residue subunit formed by domains arranged in the order of D9-D3-A1-A2-A3-D4-B-C. 5 The A1 domain contains the binding site for the platelet receptor GPIba 6 ; the cleavage site for the enzyme ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs)-13 is localized in the A2 domain, 7 and the A3 domain binds to collagen. 8 Unlike the A3 domain, both the A1 and A2 domains do not have access to their ligands until their domain structure is altered.…”

Section: Introductionmentioning

confidence: 99%

Platelet adhesion involves a novel interaction between vimentin and von Willebrand factor under high shear stress

Behymer

Correa

et al. 2014

Blood

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• Vimentin expressed on the platelet surface serves as adhesive receptor for VWF.The interaction between platelet receptor glycoprotein Iba and the A1 domain of von Willebrand factor (VWF) mediates tethering/translocation of platelets to sites of vascular injury. Unexpectedly, we observed platelets translocating over A1A2A3 domains protein slower than on A1 domain at high shear stress. This observation suggests an additional interaction between A domains and an adhesive receptor. We investigated vimentin because we have data showing the interaction of vimentin with the A2 domain of VWF. Moreover, vimentin is expressed on the platelet surface. This novel interaction was analyzed by using purified VWF, recombinant proteins, anti-vimentin antibodies, parallel flow chamber adhesion assays, flow cytometry, and vimentin-deficient murine platelets. The active form of VWF bound to vimentin, and the purified A2 domain blocked that binding. The interaction of a gain-of-function A1A2A3 mutant with platelet was reduced using anti-vimentin antibody. Platelet adhesion to wild-type (WT) A1A2A3 protein, collagen, and fibrin(ogen) was inhibited (32-75%) by anti-vimentin antibody under high shear stress. Compared with WT mice, platelets from vimentin-deficient mice had a reduced flow-dependent adhesion to both collagen and purified murine VWF. Last, the vimentin knockout mice had a prolonged tail bleeding time. The results describe that platelet vimentin engages VWF during platelet adhesion under high shear stress. (Blood. 2014;123(17):2715-2721 Introduction Atherothrombotic events, including acute coronary syndrome and stroke, are the result of platelet adhesion and activation on the ruptured atherosclerotic plaques. This platelet-mediated arterial thrombosis starts with the contact of the rapidly flowing platelets to components of the damaged blood vessel. von Willebrand factor (VWF), a multimeric plasma and subendothelial glycoprotein, is relevant in mediating platelet adhesion and activation at sites of lesions in the coronary arteries, where high shear conditions prevail.1,2 VWF captures the circulating platelets through its interaction with the platelet receptor glycoprotein (GP)Ib/IX/V complex. This interaction is responsible for the tethering, rolling, and activation of platelets that eventually become firmly adhered, leading to thrombus formation within a coronary artery. 3,4 Mature VWF consists of a 2050-residue subunit formed by domains arranged in the order of D9-D3-A1-A2-A3-D4-B-C. 5 The A1 domain contains the binding site for the platelet receptor GPIba 6 ; the cleavage site for the enzyme ADAMTS (a disintegrin and metalloproteinase with thrombospondin motifs)-13 is localized in the A2 domain, 7 and the A3 domain binds to collagen. 8 Unlike the A3 domain, both the A1 and A2 domains do not have access to their ligands until their domain structure is altered.9 This structural modification can be induced by mutations, 10 hydrodynamic forces, 11 immobilization on a surface, or artificially with the modulator ristocetin....

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von Willebrand Factor without the A2 Domain Is Resistant to Proteolysis

Cited by 30 publications

References 28 publications

An Experimental Model to Study the in Vivo Survival of von Willebrand Factor

An Experimental Model to Study the in Vivo Survival of von Willebrand Factor

P-selectin glycoprotein ligand 1 and β2-integrins cooperate in the adhesion of leukocytes to von Willebrand factor

Platelet adhesion involves a novel interaction between vimentin and von Willebrand factor under high shear stress

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