SEARCH:   Advanced

Blood, 1 December 2006, Vol. 108, No. 12, pp. 3753-3756. Prepublished online as a Blood First Edition Paper on August 15, 2006; DOI 10.1182/blood-2006-03-011965. This Article Abstract Full Text (PDF) Supplemental Tables and Figure All Versions of this Article: blood-2006-03-011965v1 108/12/3753    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Lisman, T. Articles by Farndale, R. W. Search for Related Content PubMed PubMed Citation Articles by Lisman, T. Articles by Farndale, R. W. Related Collections Hemostasis, Thrombosis, and Vascular Biology Cell Adhesion and Motility Free Research Articles Related Article in Blood Online Social Bookmarking                   What's this?  Previous Article  |  Table of Contents  |  Next Article  HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY Brief report A single high-affinity binding site for von Willebrand factor in collagen III, identified using synthetic triple-helical peptides Ton Lisman, Nicolas Raynal, Dafna Groeneveld, Ben Maddox, Anthony R. Peachey, Eric G. Huizinga, Philip G. de Groot, and Richard W. Farndale From the Department of Clinical Chemistry and Haematology, University Medical Centre Utrecht, The Netherlands; the Department of Biochemistry, University of Cambridge, United Kingdom; and the Department of Crystal and Structural Chemistry, Utrecht University, The Netherlands.    Abstract Top Abstract Introduction Materials and methods Results and discussion Authorship References   The essential event in platelet adhesion to the injured blood vessel wall is the binding to subendothelial collagen of plasma von Willebrand factor (VWF), a protein that interacts transiently with platelet glycoprotein Ib (GPIb), slowing circulating platelets to facilitate firm adhesion through collagen receptors, including integrin 21 and GpVI. To locate the site in collagen that binds VWF, we synthesized 57 overlapping triple-helical peptides comprising the whole triple-helical domain of collagen III. Peptide no. 23 alone bound VWF, with similar affinity to that of native collagen III. Immobilized peptide no. 23 supported platelet adhesion under static and flow conditions, processes blocked by an antibody that prevents collagen from binding the VWF A3 domain. Truncated and alanine-substituted peptides derived from no. 23 either strongly interacted with both VWF and platelets or lacked both VWF and platelet binding. Thus, we identified the sequence RGQOGVMGF (O is hydroxyproline) as the minimal VWF-binding sequence in collagen III.    Introduction Top Abstract Introduction Materials and methods Results and discussion Authorship References   The interaction of collagen with von Willebrand factor (VWF) requires unique structural properties in both proteins. Optimal hemostatic function requires multimerization of up to 50 VWF monomers in circulating plasma; higher-order multimers bind collagen more tightly than smaller assemblies of VWF.1 Several collagens occur in the vessel wall, of which collagens I and III are considered most important in supporting platelet adhesion to the damaged vasculature.2 We have identified the residues in the VWF A3 domain that bind collagen III, using site-directed mutagenesis guided by the crystal structure of the VWF A3 domain in complex with a monoclonal antibody (RU5) that inhibits its interaction with collagen.3,4 Nishida et al mapped the collagen-binding mode of the A3 domain by nuclear magnetic resonance and confirmed results by site-directed mutagenesis.5 However, the VWF-binding site(s) in collagen is unknown, although progress in understanding how collagen interacts with integrin 21 and GpVI has been made using short synthetic triple-helical peptide analogues of collagen,6,7 including the Collagen III Toolkit.8 We used the same approach to identify the high-affinity VWF-binding site in human collagen III, information that may help to develop the collagen-VWF interaction as an antithrombotic target.9,10    Materials and methods Top Abstract Introduction Materials and methods Results and discussion Authorship References   Peptide synthesis The synthesis and characterization of the 57 overlapping triple-helical peptides of the Collagen III Toolkit (Table S1, available at the Blood website; see the Supplemental Materials link at the top of the online article) is detailed elsewhere.8 The same approach was used to synthesize and verify derivative peptides (Table S2). The sequence of peptide no. 23 is GPC-(GPP)5-GPOGPSGPRGQOGVMGFOGPKGNDGAO-(GPP)5-GPC-NH2, and of the minimal VWF-binding derivative peptide, GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2. Static platelet-binding assay Blood was obtained from the antecubital vein of informed volunteers, in accordance with the Helsinki protocol, into 0.105-M citrate Vacutainers (Becton Dickinson, Oxford, United Kingdom). Platelet-rich plasma was prepared after 2 spins for 1 minute at 1200g. Then, 10% (vol/vol) of ACD buffer (39 mM citric acid, 75 mM trisodium citrate, 135 mM D-glucose, pH 4.5) and prostaglandin E1 (280-nM final concentration) were added, and the platelets were pelleted for 12 minutes at 700g, then resuspended in 6 mL buffer (5.5 mM D-glucose, 128 mM NaCl, 4.26 mM Na2HPO4, 7.46 mM NaH2PO4, 4.77 mM trisodium citrate, 2.35 mM citric acid, 0.35% bovine serum albumin [BSA], pH 6.5). Prostaglandin E1 was added similarly, and the platelets were spun for 6 minutes at 700g. Platelets were resuspended to 1.25 x 108 platelets/mL in adhesion buffer (0.05 M Tris-HCl, 0.14 M NaCl, 0.1% BSA, pH 7.4), and the adhesion of platelets from 100-µL portions was determined colorimetrically, as described.11 Peptides (1 µg/well) were coated onto Immulon-2-HB plates (Nunc, Rochester, NY), conditions supporting maximal platelet binding to peptide no. 23. The IIb3 antagonist, GR144053F (2 µM; Calbiochem, Nottingham, United Kingdom), or antibody RU5 (2 µg/mL) was preincubated for 15 minutes with platelets, where indicated. Approval for these studies was obtained from the University of Cambridge Human Biology Research Ethics Committee. Solid-phase binding assays To measure binding of VWF12 (purified from Haemate P; Aventis-Behring, Hattersheim am Main, Germany), 96-well plates were coated with peptides (1 µg/well) or collagen III (10 µg/well; Sigma, St Louis, MO) and incubated with VWF (1 µg/mL), which was detected with a polyclonal antibody.12 View larger version (17K): [in this window] [in a new window]   Figure 1.. Identification of a single peptide containing 27 amino acids of collagen III sequence that specifically binds VWF. (A) The peptides (10 µg/mL) of the Collagen III Toolkit (Table S1) were each immobilized on a 96-well plate, and the adhesion of plasma-derived VWF (1 µg/mL) was determined. (B) Immobilized peptide no. 23 was incubated with plasma-derived VWF (1 µg/mL) in presence or absence of monoclonal antibody RU5 (1 µg/mL), or with recombinant wild-type, delta-A3, or His1023Ala VWF (all at 1 µg/mL). In panels A-B, bound VWF was detected as described in "Solid-phase binding assays," and the mean ± SD is shown from a representative of 3 independent experiments, each performed in duplicate. (C) Immobilized peptide no. 23 or immobilized collagen III (100 µg/mL) was incubated with increasing concentrations of VWF, detected as in panel A. (D) Peptide no. 23 or vehicle was coated in a 96-well plate (10 µg/mL) or sprayed onto a coverslip (0.5 µg/cm2) and incubated with washed platelets or perfused with whole blood for 5 minutes at a shear rate of 300 s–1 in presence or absence of RU5 (1 µg/mL). Uncoated wells or coverslips served as controls. The adhesion of platelets bound in the static assay () and the surface coverage of platelets () were each determined as described in "Static platelet-binding assay" and "Solid-phase binding assays." Shown is the mean ± SD of 3 independent experiments each performed in triplicate.   Platelet binding under flow conditions Peptides (0.5 µg/cm2) or collagen III (6.5 µg/cm2) was spray-coated onto Thermanox coverslips (Nunc), then blocked with 1% human albumin in PBS and perfused at 22°C with citrated whole blood for 5 minutes at a shear rate of 300 s–1 using a single-pass perfusion chamber,13 and platelet adhesion was evaluated.14 The institutional review board of the University Medical Centre, University of Utrecht, approved these studies. Surface plasmon resonance A Biacore 2000 system (Biacore AB, Uppsala, Sweden) was used for surface plasmon resonance analysis (SPR). Peptides were immobilized on a CM5 sensor chip via the cysteine-free thiols in the peptide N- and C-termini, using the manufacturer's procedures. Binding to a channel coated with control peptide, GPC-(GPP)10-GPC was used to correct the binding of VWF or A3 domain to collagen- or other peptide-coated channels, and VWF concentration used to calculate affinities was based on that of the monomer, measured by enzyme-linked immunosorbent assay.    Results and discussion Top Abstract Introduction Materials and methods Results and discussion Authorship References   Mapping the VWF-binding site within collagen III A single Collagen III Toolkit peptide, no. 23, bound plasma-derived human VWF (Figure 1A), a process that resembled VWF binding to collagen: (1) the interaction was abolished by a monoclonal antibody, RU5,3 directed against the collagen-binding site of VWF A3 domain (Figure 1B); (2) dysfunctional recombinant variants of VWF, delta A3 VWF15 (which lacks the A3 domain) and His1023Ala VWF3 (in which a crucial A3 amino acid is substituted), showed severely reduced binding to peptide no. 23 (Figure 1B); (3) VWF had similar affinity for peptide no. 23 as for full-length collagen III measured in a solid-phase assay (Figure 1C); and (4) peptide no. 23 bound washed human platelets in static assays, most likely reflecting the presence on the platelet surface of residual plasma-derived VWF or VWF inevitably secreted during platelet preparation, and supported platelet deposition from flowing whole blood at low shear rate (300 s–1; Figure 1D). The high density (20-fold that of native collagen) of VWF-binding sites on the peptide-coated surface may be responsible for the static binding activity, and the complete blockade of these events by RU5 shows the fundamental role of VWF. Under static conditions, antagonism of either integrin IIb3 or GpVI slightly impaired platelet binding to peptide no. 23, consistent with minor involvement of other receptors but with GPIb being the main VWF receptor (data not shown). A nonhelical derivative of peptide no. 23, flanked by GAP rather than GPP repeats, bound neither VWF nor platelets (data not shown), indicating the same requirement for triple-helical structure for the recognition of collagen by VWF as for 21 and GpVI.6,7 Identification of the minimal collagen sequence required for VWF binding A set of truncated triple-helical derivatives of peptide no. 23 was synthesized, including an alanine-scanned set, peptides 7 to 14 (Table S2). Peptides either bound VWF (Figure 2A) and platelets (Figure 2B) strongly or completely lacked affinity for both VWF and platelets. Thus we identified RGQOGVMGF as the minimum sequence within collagen III required to bind VWF. Residues R, O, V, and F appear crucial for VWF binding, but not Q and M. The triple-helical peptide GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2, its parent peptide no. 23, and full-length collagen bound VWF in solid-phase assays with similar affinity (data not shown). SPR allowed more detailed kinetic analysis. A recombinant VWF A3 domain16 bound the immobilized peptides with modest affinity (Kd, 1.8 µM for peptide no. 23; Kd, 2.5 µM for GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2; data not shown), whereas full-length, plasma-derived VWF displayed much higher affinity, attributable to its multimeric nature (Kd, 2.1 nM for peptide no. 23; Kd, 2.5 nM for GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2; Figure 2C). These affinity constants are consistent with those observed for A3 and VWF binding to full-length collagen12,17 and imply a single VWF-binding site within collagen III. View larger version (32K): [in this window] [in a new window]   Figure 2.. Identification of the minimal collagen sequence required for VWF binding. (A) The truncated and alanine-modified peptides derived from peptide no. 23 (Table S2) were immobilized on a 96-well plate and incubated with plasma-derived VWF (1 µg/mL), and bound VWF was detected as in Figure 1. (B) The same peptides were spray-coated onto Thermanox coverslips (0.5 µg/cm2) and perfused with whole blood at a shear rate of 300 s–1 for 5 minutes; then surface coverage was measured as described for Figure 1. (C) Peptide no. 23 and peptide GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2 were immobilized onto a Biacore CM5 sensor chip via their free cysteine residues. The equilibrium binding capacity of the peptides was determined for different concentrations of plasma-derived VWF. (D) Collagen (100 µg/mL) was immobilized on a 96-well plate or a Thermanox coverslip and incubated with purified VWF (1 µg/mL) or perfused with whole blood in the presence or absence of different concentrations of peptide GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2. VWF binding and platelet deposition were evaluated as in Figure 1. Shown is a representative of 3 independent experiments, each performed in duplicate (A), triplicate (B,D), or as single determinations (C). Error bars denote SD.   We previously reported putative VWF-binding sites in collagen III,18 but VWF bound weakly to these synthetic triple-helical peptides compared with full-length collagen. These peptides were short stretches of cyanogen bromide fragments of bovine collagen, and so lacked the intact high-affinity binding site, which was cleaved in the parent collagen by cyanogen bromide digestion at the methionine residue within the sequence that we now report. The triple-helical peptide GPC-(GPP)5-GPRGQOGVMGFO-(GPP)5-GPC-NH2 at 500 µg/mL ( 40 µM) inhibited binding of VWF, or platelets from whole blood, to immobilized collagen III under flow conditions almost completely (Figure 2D). These results confirm the sequence RGQOGVMGF as the major, high-affinity VWF-binding site and exclude the prominent role for the VWF A1 domain in collagen binding postulated previously.19 We cannot exclude the possibility that the residual binding of the A3-domain mutant, His1023Ala, reflects weak binding of VWF to peptide no. 23 through A1, nor that binding of A3 is required to render A1 competent to bind collagen at a separate site. Our data suggest that inhibition of VWF binding to collagen, by peptides, antibodies, or other antagonists that target this nonapeptide collagen sequence, may be a useful therapeutic strategy. Current antiplatelet therapy addresses platelet activation through amplification pathways including purinergic receptors and thromboxane generation, or the up-regulation of the fibrinogen receptor integrin IIb3. Previous use of in vivo models suggested that inhibition of the collagen-VWF interaction with a monoclonal antibody or a naturally occurring collagen-binding protein, by targeting primary platelet adhesion to the arterial subendothelium, has a better risk-benefit ratio than inhibition of platelet-platelet interaction,9,10 and might provide a valuable alternative to the antiplatelet therapies in current clinical practice (recently reviewed in Ahrens et al20). The sequence RGQOGVMGF (underlined) occurs in only one other human protein, collagen II. A peptide, GPC-(GPP)5-GAOGEDGROGPOGPQGARGQOGVMGFO-(GPP)5-GPC (amino acids 510-536 from collagen II) bound VWF with high affinity (data not shown). This may be irrelevant to hemostasis, since collagen II is not found in vascular subendothelium, being largely restricted to cartilage and vitreous humor. The sequence RGQOGVMGF is 100% conserved in collagen III from mouse, rat, cow, and chicken. In human collagen I, a heterotrimer comprising 2 1 and 1 2 chains, a closely related sequence occurs in the 1 chain, differing by a single amino acid (RGQAGVMGF). This O-to-A substitution prevented the VWF-binding activity in our homotrimeric synthetic peptide from the Ala-scanned set. In this region, the sequence RGQAGVMGF of the human 1(I) chain aligns with the sequence RGEOGNIGF of the 2(I) chain, suggesting that the essential O in position 4 of the VWF-binding homotrimer, although substituted with an A in the 1(I) chain, is provided by position 4 of the 2(I) chain sequence, so that VWF may bind collagens I, II, and III in an identical manner. Modeling experiments, shown in Figure S1, support this conclusion.    Authorship Top Abstract Introduction Materials and methods Results and discussion Authorship References   The authors declare no competing financial interests. T.L., N.R., P.G.G., and R.W.F. contributed equally to this work.    Acknowledgements   This work was supported in part by a grant from the Wellcome Trust.    Footnotes   Submitted March 24, 2006; accepted July 24, 2006. Prepublished online as Blood First Edition Paper, August 15, 2006; DOI 10.1182/blood-2006-03-011965. The online version of this article contains a data supplement. An Inside Blood analysis of this article appears at the front of this issue. The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 USC section 1734. Correspondence: Richard W. Farndale, Department of Biochemistry, Downing Site, Cambridge CB2 1QW, United Kingdom; e-mail: rwf10{at}cam.ac.uk.    References Top Abstract Introduction Materials and methods Results and discussion Authorship References   Santoro SA. Preferential binding of high molecular weight forms of von Willebrand factor to fibrillar collagen. Biochim Biophys Acta. 1983;756: 123-126.[Medline] [Order article via Infotrieve] Farndale RW, Sixma JJ, Barnes MJ, de Groot PG. Role of collagen in thrombosis and haemostasis. J Thromb Haemost. 2004;2: 561-573.[CrossRef][Medline] [Order article via Infotrieve] Romijn RA, Bouma B, Wuyster W, et al. Identification of the collagen-binding site of the von Willebrand factor A3-domain. J Biol Chem. 2001; 276: 9985-9991.[Abstract/Free Full Text] Romijn RA, Westein E, Bouma B, et al. Mapping the collagen-binding site in the von Willebrand factor-A3 domain. J Biol Chem. 2003;278: 15035-15039.[Abstract/Free Full Text] Nishida N, Sumikawa H, Sakakura M, et al. Collagen-binding mode of vWF-A3 domain determined by a transferred cross-saturation experiment. Nat Struct Biol. 2003;10: 53-58.[CrossRef][Medline] [Order article via Infotrieve] Knight CG, Morton LF, Onley DJ, et al. Collagen-platelet interaction: Gly-Pro-Hyp is uniquely specific for platelet GpVI and mediates platelet activation by collagen. Cardiovasc Res. 1999;41: 450-457.[Abstract/Free Full Text] Emsley J, Knight CG, Farndale RW, Barnes MJ, Liddington RC. Structural basis of collagen recognition by 21. Cell. 2000;101: 47-56.[CrossRef][Medline] [Order article via Infotrieve] Raynal N, Hamaia SW, Siljander PR, et al. Use of synthetic peptides to locate novel integrin alpha2beta1-binding motifs in human collagen III. J Biol Chem. 2006;281: 3821-3831.[Abstract/Free Full Text] Wu D, Vanhoorelbeke K, Cauwenberghs N, et al. Inhibition of the von Willebrand (VWF)-collagen interaction by an antihuman VWF monoclonal antibody results in abolition of in vivo arterial platelet thrombus formation in baboons. Blood. 2002;99: 3623-3628.[Abstract/Free Full Text] Lasser G, Guchhait P, Ellsworth JL, et al. C1qTNF-related protein-1 (CTRP-1): a vascular wall protein that inhibits collagen-induced platelet aggregation by blocking VWF binding to collagen. Blood. 2006;107: 423-430.[Abstract/Free Full Text] Onley DJ, Knight CG, Tuckwell DS, Barnes MJ, Farndale RW. Micromolar Ca2+ is essential for Mg2+-dependent binding of collagen by the integrin 21 in human platelets. J Biol Chem. 2000; 275: 24560-24566.[Abstract/Free Full Text] van der Plas RM, Gomes L, Marquart JA, et al. Binding of von Willebrand factor to collagen type III: role of specific amino acids in the collagen binding domain of vWF and effects of neighboring domains. Thromb Haemost. 2000;84: 1005-1011.[Medline] [Order article via Infotrieve] Sixma JJ, de Groot PG, van Zanten H, Ijsseldijk M. A new perfusion chamber to detect platelet adhesion using a small volume of blood. Thromb Res. 1998;92: S43-S46.[CrossRef][Medline] [Order article via Infotrieve] Lisman T, Moschatsis S, Adelmeijer J, Nieuwenhuis HK, De Groot PG. Recombinant factor VIIa enhances deposition of platelets with congenital or acquired alpha IIb beta 3 deficiency to endothelial cell matrix and collagen under conditions of flow via tissue factor-independent thrombin generation. Blood. 2003; 101: 1864-1870.[Abstract/Free Full Text] Lankhof H, van Hoeij M, Schiphorst ME, et al. A3 domain is essential for interaction of von Willebrand factor with collagen type III. Thromb Haemost. 1996;75: 950-958.[Medline] [Order article via Infotrieve] Huizinga EG, van der Plas RM, Kroon J, Sixma JJ, Gros P. Crystal structure of the A3 domain of human von Willebrand factor: implications for collagen binding. Structure. 1997;5: 1147-1156.[Medline] [Order article via Infotrieve] Cruz MA, Yuan H, Lee JR, Wise RJ, Handin RI. Interaction of the von Willebrand factor (vWF) with collagen: localization of the primary collagen-binding site by analysis of recombinant vWF a domain polypeptides. J Biol Chem. 1995;270: 10822-10827.[Abstract/Free Full Text] Verkleij MW, Ijsseldijk MJW, Heijnen-Snyder GJ, et al. Adhesive domains in the collagen III fragment 1(III)CB4 that support 21- and von Willebrand factor-mediated platelet adhesion under flow conditions. Thromb Haemost. 1999;82: 1137-1144.[Medline] [Order article via Infotrieve] Morales LD, Martin C, Cruz MA. The interaction of von Willebrand factor-A1 domain with collagen: mutation G1324S (type 2M von Willebrand disease) impairs the conformational change in A1 domain induced by collagen. J Thromb Haemost. 2006;4: 417-425.[CrossRef][Medline] [Order article via Infotrieve] Ahrens I, Bode C, Peter K. Inhibition of platelet activation and aggregation. Handb Exp Pharmacol. 2005;170: 443-462. CiteULike    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this? Related Article in Blood Online: When collagen meets VWF Hans Deckmyn and Karen Vanhoorelbeke Blood 2006 108: 3628. [Full Text] [PDF] This article has been cited by other articles: A. B. Herr and R. W. Farndale Structural Insights into the Interactions between Platelet Receptors and Fibrillar Collagen J. Biol. Chem., July 24, 2009; 284(30): 19781 - 19785. [Abstract] [Full Text] [PDF] E. Hohenester, T. Sasaki, C. Giudici, R. W. Farndale, and H. P. Bachinger Structural basis of sequence-specific collagen recognition by SPARC PNAS, November 25, 2008; 105(47): 18273 - 18277. [Abstract] [Full Text] [PDF] S. M. Sweeney, J. P. Orgel, A. Fertala, J. D. McAuliffe, K. R. Turner, G. A. Di Lullo, S. Chen, O. Antipova, S. Perumal, L. Ala-Kokko, et al. Candidate Cell and Matrix Interaction Domains on the Collagen Fibril, the Predominant Protein of Vertebrates J. Biol. Chem., July 25, 2008; 283(30): 21187 - 21197. [Abstract] [Full Text] [PDF] C. Giudici, N. Raynal, H. Wiedemann, W. A. Cabral, J. C. Marini, R. Timpl, H. P. Bachinger, R. W. Farndale, T. Sasaki, and R. Tenni Mapping of SPARC/BM-40/Osteonectin-binding Sites on Fibrillar Collagens J. Biol. Chem., July 11, 2008; 283(28): 19551 - 19560. [Abstract] [Full Text] [PDF] G. E. Jarvis, N. Raynal, J. P. Langford, D. J. Onley, A. Andrews, P. A. Smethurst, and R. W. Farndale Identification of a major GpVI-binding locus in human type III collagen Blood, May 15, 2008; 111(10): 4986 - 4996. [Abstract] [Full Text] [PDF] A. D. Konitsiotis, N. Raynal, D. Bihan, E. Hohenester, R. W. Farndale, and B. Leitinger Characterization of High Affinity Binding Motifs for the Discoidin Domain Receptor DDR2 in Collagen J. Biol. Chem., March 14, 2008; 283(11): 6861 - 6868. [Abstract] [Full Text] [PDF] E. Calvo, F. Tokumasu, O. Marinotti, J.-L. Villeval, J. M. C. Ribeiro, and I. M. B. Francischetti Aegyptin, a Novel Mosquito Salivary Gland Protein, Specifically Binds to Collagen and Prevents Its Interaction with Platelet Glycoprotein VI, Integrin {alpha}2beta1, and von Willebrand Factor J. Biol. Chem., September 14, 2007; 282(37): 26928 - 26938. [Abstract] [Full Text] [PDF] C. V. Denis and D. D. Wagner Platelet Adhesion Receptors and Their Ligands in Mouse Models of Thrombosis Arterioscler Thromb Vasc Biol, April 1, 2007; 27(4): 728 - 739. [Abstract] [Full Text] [PDF] This Article Abstract Full Text (PDF) Supplemental Tables and Figure All Versions of this Article: blood-2006-03-011965v1 108/12/3753    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Lisman, T. Articles by Farndale, R. W. Search for Related Content PubMed PubMed Citation Articles by Lisman, T. Articles by Farndale, R. W. Related Collections Hemostasis, Thrombosis, and Vascular Biology Cell Adhesion and Motility Free Research Articles Related Article in Blood Online Social Bookmarking                   What's this?

	Copyright © 2006 by American Society of Hematology         Online ISSN: 1528-0020