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Thromb Haemost 1999; 82(03): 1160-1163
DOI: 10.1055/s-0037-1614346
Letters to the Editor
Schattauer GmbH

The Collagen-binding Leech Products rLAPP and Calin Prevent both von Willebrand Factor and α2β1 (GPIa/IIa)-I-domain Binding to Collagen in a Different Manner

H. Depraetere
1   From the Laboratory for Thrombosis Research, IRC, KU Leuven Campus Kortrijk, Belgium
,
A. Kerekes   **
1   From the Laboratory for Thrombosis Research, IRC, KU Leuven Campus Kortrijk, Belgium
,
H. Deckmyn
1   From the Laboratory for Thrombosis Research, IRC, KU Leuven Campus Kortrijk, Belgium
› Author Affiliations
Further Information

Publication History

Received 01 January 1999

Accepted 07 May 1999

Publication Date:
09 December 2017 (online)

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Summary

Calin and rLAPP are two natural inhibitors that are able to inhibit the vWF-binding and platelet adhesion to collagen both under static and flow conditions. In this study we demonstrate that both rLAPP and Calin prevent α2I-domain binding to human collagen type I with an IC50 of 5 μg/ml. However, although both vWF and α2I-domain binding to collagen is prevented by rLAPP and Calin, the latter two do not bind to the same collagen site since Calin only partially could compete with rLAPP for binding to collagen. Also vWF and the α2I-domain were unable to compete completely with each other for the binding to collagen. So the following hypothesis can be made: the binding sites of vWF and of the α2I-domain on human collagen type I are different but close to each other since rLAPP could inhibit both interactions, and thus should bind to an overlapping epitope. The Calin preparation on the other hand may still contain two active principles, one interfering with vWF-binding, the other with the α2I-domain-binding to collagen.

** Present address: Dr. A. Kerekes, Department of Clinical Chemistry, University of Debrecen Medical School, P.O. Box 40, H-4012 Debrecen, Hungary


 

  • References

  • 1 Santoro SA. Identification of a 160,000 dalton platelet membrane protein that mediates the initial divalent cation-dependent adhesion of platelets to collagen. Cell 1986; 46: 913-20.
    CrossrefPubMedGoogle Scholar
  • 2 Nieuwenhuis HK, Akkerman JW, Houdijk WP, Sixma JJ. Human blood platelets showing no response to collagen fail to express surface glycoprotein Ia. Nature 1985; 318: 470-2.
    CrossrefPubMedGoogle Scholar
  • 3 Santoro SA, Zutter MM. The alpha-2 beta-1 integrin: a collagen receptor on platelets and other cells. Thromb Haemost 1995; 74: 813-21.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 4 Kamata T, Puzon W, Takada Y. Identification of putative ligand binding sites within I domain of integrin alpha 2 beta 1 (VLA-2, CD49b/CD29). J Biol Chem 1994; 269: 9659-63.
    PubMedGoogle Scholar
  • 5 Kamata T, Takada Y. Direct binding of collagen to the I domain of integrin alpha 2 beta 1 (VLA-2, CD49b/CD29) in a divalent cation-independent manner. J Biol Chem 1994; 269: 26006-10.
    PubMedGoogle Scholar
  • 6 Bahou WF, Potter CL, Mirza H. The VLA-2 (alpha 2 beta 1) I domain functions as a ligand-specific recognition sequence for endothelial cell attachment and spreading: molecular and functional characterization. Blood 1994; 84: 3734-41.
    PubMedGoogle Scholar
  • 7 Depraetere H, Wille C, Gansemans Y, Stanssens P, Lauwereys M, Baruch D, De Reys S, Deckmyn H. The integrin alpha 2 beta 1 (GPIa/IIa)-I-domain inhibits platelet-collagen interaction. Thromb Haemost 1997; 77: 981-5.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 8 Kehrel B, Wierwille S, Clemetson KJ, Anders O, Steiner M, Knight CG, Farndale RW, Okuma M, Barnes MJ. Glycoprotein VI is a major collagen receptor for platelet activation: it recognizes the platelet-activating quaternary structure of collagen, whereas CD36, glycoprotein IIb/IIIa, and von Willebrand factor do not. Blood 1998; 91: 491-9.
    PubMedGoogle Scholar
  • 9 Gibbins JM, Okuma M, Farndale R, Barnes M, Watson SP. Glycoprotein VI is the collagen receptor in platelets which underlies tyrosine phosphorylation of the Fc receptor gamma-chain. FEBS Lett 1997; 413: 255-9.
    CrossrefPubMedGoogle Scholar
  • 10 Diaz Ricart M, Tandon NN, Carretero M, Ordinas A, Bastida E, Jamieson GA. Platelets lacking functional CD36 (glycoprotein IV) show reduced adhesion to collagen in flowing whole blood. Blood 1993; 82: 491-6.
    PubMedGoogle Scholar
  • 11 Deckmyn H, Van Houtte E, Vermylen J. Disturbed platelet aggregation to collagen associated with an antibody against an 85- to 90-Kd platelet glycoprotein in a patient with prolonged bleeding time. Blood 1992; 79: 1466-71.
    PubMedGoogle Scholar
  • 12 Chiang TM, Rinaldy A, Kang AH. Cloning, characterization, and functional studies of a nonintegrin platelet receptor for type I collagen. J Clin Invest 1997; 100: 514-21.
    CrossrefPubMedGoogle Scholar
  • 13 Savage B, Almus-Jacobs F, Ruggeri ZM. Specific synergy of multiple substrate-receptor interactions in platelet thrombus formation under flow. Cell 1998; 94: 657-66.
    CrossrefPubMedGoogle Scholar
  • 14 Connolly TM, Jacobs JW, Condra C. An inhibitor of collagen-stimulated platelet activation from the salivary glands of the Haementeria officinalis leech. I. Identification, isolation, and characterization. J Biol Chem 1992; 267: 6893-8.
    PubMedGoogle Scholar
  • 15 Keller PM, Schultz LD, Condra C, Karczewski J, Connolly TM. An inhibitor of collagen-stimulated platelet activation from the salivary glands of the Haementeria officinalis leech. II. Cloning of the cDNA and expression. J Biol Chem 1992; 267: 6899-904.
    PubMedGoogle Scholar
  • 16 van Zanten GH, Connolly TM, Schiphorst ME, de Graaf S, Slootweg PJ, Sixma JJ. Recombinant leech antiplatelet protein specifically blocks platelet deposition on collagen surfaces under flow conditions. Arterioscler Thromb Vasc Biol 1995; 15: 1424-31.
    CrossrefPubMedGoogle Scholar
  • 17 Harsfalvi J, Stassen JM, Hoylaerts MF, Van Houtte E, Sawyer RT, Vermylen J, Deckmyn H. Calin from Hirudo medicinalis, an inhibitor of von Willebrand factor binding to collagen under static and flow conditions. Blood 1995; 85: 705-11.
    PubMedGoogle Scholar
  • 18 Munro R, Jones CP, Sawyer RT. Calin – a platelet adhesion inhibitor from the saliva of the medicinal leech. Blood Coagul Fibrinolysis 1991; 2: 179-84.
    CrossrefPubMedGoogle Scholar
  • 19 Deckmyn H, Stassen JM, Vreys I, Van Houtte E, Sawyer RT, Vermylen J. Calin from Hirudo medicinalis, an inhibitor of platelet adhesion to collagen, prevents platelet-rich thrombosis in hamsters. Blood 1995; 85: 712-9.
    PubMedGoogle Scholar
  • 20 Saelman EU, Nieuwenhuis HK, Hese KM, de Groot PG, Heijnen HF, Sage EH, Williams S, McKeown L, Gralnick HR, Sixma JJ. Platelet adhesion to collagen types I through VIII under conditions of stasis and flow is mediated by GPIa/IIa (alpha 2 beta 1-integrin). Blood 1994; 83: 1244-50.
    PubMedGoogle Scholar
  • 21 Schaffer LW, Davidson JT, Siegl PK, Gould RJ, Nutt RF, Brady SF, Connolly TM. Recombinant leech antiplatelet protein prevents collagen-mediated platelet aggregation but not collagen graft thrombosis in baboons. Arterioscler Thromb 1993; 13: 1593-601.
    CrossrefPubMedGoogle Scholar
  • 22 Knight CG, Morton LF, Onley DJ, Peachey AR, Messent AJ, Smethurst PA, Tuckwell DS, Farndale RW, Barnes MJ. Identification in collagen type I of an integrin alpha2betal-binding site containing an essential GER sequence. J Biol Chem 1998; 273: 33287-94.
    CrossrefPubMedGoogle Scholar