Identification of a VWF peptide antagonist that blocks platelet adhesion under high shear conditions by selectively inhibiting the VWF-collagen interaction

T. Szanto, K. Vanhoorelbeke, G. Toth, A. Vandenbulcke, J. Toth, W. Noppe, H. Deckmyn, J. Hársfalvi

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background: Because the collagen-VWF-GPIb/IX/V axis plays an important role in thrombus formation, it represents a promising target for development of new antithrombotic agents. Objectives: We used phage display to identify potential small peptides that interfere with the VWF-collagen binding and might serve as lead products for the development of possible oral antithrombotic compounds. Methods: A random linear heptamer peptide library was used to select VWF-binding peptides. Results: We identified a phage clone, displaying the YDPWTPS sequence, further referred to as L7-phage, that bound to VWF in a specific and a dose-dependent manner. This L7-phage specifically inhibited the VWF-collagen interaction under both static and flow conditions. Epitope mapping using deletion mutants of VWF revealed that the L7-phage does not bind to the known collagen-binding A3 domain within VWF, but to the more carboxyterminal situated C domain. This inhibition was not due to steric hindrance of the A3 domain-collagen interaction by the L7-phage. Indeed, a tetrabranched multi-antigen peptide (MAP) presenting four copies of the peptide, but not the scrambled MAP, also inhibited VWF-collagen interaction under conditions of high shear stress at a concentration of 148 nmol L-1. Conclusions: Based on these results, we conclude that we have identified the first peptide antagonist that binds to the VWF C domain and by this specifically inhibits the VWF binding to collagen, suppressing platelet adhesion and aggregation under high shear conditions. As a consequence, this peptide and its future derivates are potentially interesting antithrombotic agents.

Original languageEnglish
Pages (from-to)1680-1687
Number of pages8
JournalJournal of Thrombosis and Haemostasis
Volume7
Issue number10
DOIs
Publication statusPublished - Oct 2009

Fingerprint

Platelet Aggregation Inhibitors
Bacteriophages
Collagen
Peptides
Fibrinolytic Agents
Epitope Mapping
Antigens
Peptide Library
Platelet Aggregation
Thrombosis
Clone Cells

Keywords

  • Antithrombotic therapy
  • Collagen
  • Peptide inhibitor
  • Phage display
  • Platelet adhesion
  • Von Willebrand factor

ASJC Scopus subject areas

  • Hematology

Cite this

Identification of a VWF peptide antagonist that blocks platelet adhesion under high shear conditions by selectively inhibiting the VWF-collagen interaction. / Szanto, T.; Vanhoorelbeke, K.; Toth, G.; Vandenbulcke, A.; Toth, J.; Noppe, W.; Deckmyn, H.; Hársfalvi, J.

In: Journal of Thrombosis and Haemostasis, Vol. 7, No. 10, 10.2009, p. 1680-1687.

Research output: Contribution to journalArticle

Szanto, T. ; Vanhoorelbeke, K. ; Toth, G. ; Vandenbulcke, A. ; Toth, J. ; Noppe, W. ; Deckmyn, H. ; Hársfalvi, J. / Identification of a VWF peptide antagonist that blocks platelet adhesion under high shear conditions by selectively inhibiting the VWF-collagen interaction. In: Journal of Thrombosis and Haemostasis. 2009 ; Vol. 7, No. 10. pp. 1680-1687.
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AU - Szanto, T.

AU - Vanhoorelbeke, K.

AU - Toth, G.

AU - Vandenbulcke, A.

AU - Toth, J.

AU - Noppe, W.

AU - Deckmyn, H.

AU - Hársfalvi, J.

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N2 - Background: Because the collagen-VWF-GPIb/IX/V axis plays an important role in thrombus formation, it represents a promising target for development of new antithrombotic agents. Objectives: We used phage display to identify potential small peptides that interfere with the VWF-collagen binding and might serve as lead products for the development of possible oral antithrombotic compounds. Methods: A random linear heptamer peptide library was used to select VWF-binding peptides. Results: We identified a phage clone, displaying the YDPWTPS sequence, further referred to as L7-phage, that bound to VWF in a specific and a dose-dependent manner. This L7-phage specifically inhibited the VWF-collagen interaction under both static and flow conditions. Epitope mapping using deletion mutants of VWF revealed that the L7-phage does not bind to the known collagen-binding A3 domain within VWF, but to the more carboxyterminal situated C domain. This inhibition was not due to steric hindrance of the A3 domain-collagen interaction by the L7-phage. Indeed, a tetrabranched multi-antigen peptide (MAP) presenting four copies of the peptide, but not the scrambled MAP, also inhibited VWF-collagen interaction under conditions of high shear stress at a concentration of 148 nmol L-1. Conclusions: Based on these results, we conclude that we have identified the first peptide antagonist that binds to the VWF C domain and by this specifically inhibits the VWF binding to collagen, suppressing platelet adhesion and aggregation under high shear conditions. As a consequence, this peptide and its future derivates are potentially interesting antithrombotic agents.

AB - Background: Because the collagen-VWF-GPIb/IX/V axis plays an important role in thrombus formation, it represents a promising target for development of new antithrombotic agents. Objectives: We used phage display to identify potential small peptides that interfere with the VWF-collagen binding and might serve as lead products for the development of possible oral antithrombotic compounds. Methods: A random linear heptamer peptide library was used to select VWF-binding peptides. Results: We identified a phage clone, displaying the YDPWTPS sequence, further referred to as L7-phage, that bound to VWF in a specific and a dose-dependent manner. This L7-phage specifically inhibited the VWF-collagen interaction under both static and flow conditions. Epitope mapping using deletion mutants of VWF revealed that the L7-phage does not bind to the known collagen-binding A3 domain within VWF, but to the more carboxyterminal situated C domain. This inhibition was not due to steric hindrance of the A3 domain-collagen interaction by the L7-phage. Indeed, a tetrabranched multi-antigen peptide (MAP) presenting four copies of the peptide, but not the scrambled MAP, also inhibited VWF-collagen interaction under conditions of high shear stress at a concentration of 148 nmol L-1. Conclusions: Based on these results, we conclude that we have identified the first peptide antagonist that binds to the VWF C domain and by this specifically inhibits the VWF binding to collagen, suppressing platelet adhesion and aggregation under high shear conditions. As a consequence, this peptide and its future derivates are potentially interesting antithrombotic agents.

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KW - Phage display

KW - Platelet adhesion

KW - Von Willebrand factor

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