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Thromb Haemost 1986; 56(03): 391-396
DOI: 10.1055/s-0038-1661689
Original Article
Schattauer GmbH Stuttgart

Comparison of Tryptic Fragments of von Willebrand Factor Involved in Binding to Thrombin-Activated Platelets with Fragments Involved in Ristocetin-Induced Binding and Binding to Collagen

Wim P M Houdijk
The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Jen-pierre Girma
*   The INSERM U. 143, Hôpital de Bicêtre, Paris, France
,
Jan A van Mourik
**   The Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands
,
Jan J Sixma
The Department of Haematology, University Hospital Utrecht, The Netherlands
,
Dominique Meyer
*   The INSERM U. 143, Hôpital de Bicêtre, Paris, France
› Author Affiliations
Further Information

Publication History

Received 18 July 1986

Accepted after revision 25 September 1986

Publication Date:
18 July 2018 (online)

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Summary

Previously we have studied the binding domains on von Willebrand factor (vWF) involved in ristocetin-induced binding to platelets (ristocetin binding domain, RBD) and in the binding of vWF to collagen (collagen binding domain, CBD) using tryptic fragments of 125I-labelled vWF (21, 23). We have also reported on the RBD, CBD and the domain on vWF involved in the binding to thrombin activated platelets (thrombin binding domain, TBD) using vWF-fragments prepared by digestion with staphylococcal protease V8 (25).

In the present study, we have digested 125I-vWF with TPCK-trypsin and we have performed at various times of digestion immuno-precipitation with Mab 9, the antibody inhibiting binding of vWF to thrombin activated platelets. The data were compared with the immunoprecipitation patterns simultaneously obtained with CLB-RAg 35 which inhibits binding of vWF in the presence of ristocetin and with CLB-RAg 201, which inhibits binding of vWF to collagen. At 90 min, Mab 9 and CLB-RAg 201 precipitated similar high molecular weight bands, whereas CLB-RAg 35 precipitated bands at 180 and 120 kDa. After 24 h, Mab 9 precipitated bands at 200, 155, 116 and 85 kDa; CLB-RAg 201 precipitated a band at 48 kDa and CLB-RAg 35 a band at 116 kDa. Two-dimensional electrophoresis demonstrated that the high molecular weight bands, precipitated by Mab 9 and CLB-RAg 201 at 90 min, were identical. The 116 kDa fragment recognized by CLB-RAg 35 had a different subunit composition than the 116 kDa fragment precipitated by Mab 9.

These data indicate that there is an early split in the vWF subunit which yields a large fragment, containing the CBD and the TBD, and a smaller fragment with the RBD. Later, there is a split in the large fragment and the CBD and the TBD are separated. Comparison with data obtained with Staph protease V8 indicate that the CBD is located between the RBD and TBD.

Key words

von Willebrand factor - Platelets - Ristocetin - Collagen
 

  • References

  • 1 Hoyer LW. The factor VIII complex: Structure and function. Blood 1981; 58: 1-13
    PubMedGoogle Scholar
  • 2 Counts RB, Paskell SL, Elgee SK. Disulfide bonds and the quaternary structure of factor VIII/von Willebrand factor. J Clin Invest 1978; 62: 702-709
    CrossrefPubMedGoogle Scholar
  • 3 Meyer D, Obert B, Pietu G, Lavergne JM, Zimmerman TS. Multimeric structure of factor VII/von Willebrand factor in von Willebrand’s disease. J Lab Clin Med 1980; 95: 590-602
    PubMedGoogle Scholar
  • 4 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of factor VIII/von Willebrand factor. Blood 1981; 57: 1140-1143
    PubMedGoogle Scholar
  • 5 Fowler WE, Fretto LJ, Hamilton KK, Erickson HP, McKee PA. Substructure of human von Willebrand Factor. J Clin Invest 1985; 76: 1491-1500
    CrossrefPubMedGoogle Scholar
  • 6 Loscalzo J, Handin RI. Conformational domains and structural transitions of human von Willebrand protein. Biochemistry 1984; 23: 3880-3886
    CrossrefPubMedGoogle Scholar
  • 7 Weiss HJ, Turitto VT, Baumgartner HR. Effect of shear rate on platelet interaction with subendothelium in citrated and native blood. I. Shear rate-dependent decrease of adhesion in von Willebrand’s disease and the Bernard-Soulier syndrome. J Lab Clin Med 1978; 92: 750-764
    PubMedGoogle Scholar
  • 8 Weiss HJ, Rogers J, Brand H. Defective ristocetin-induced platelet aggregation in von Willebrand’s disease and its connection by factor VIII. J Clin Invest 1973; 52: 2697-2707
    CrossrefPubMedGoogle Scholar
  • 9 Coller BS, Peerschke EI, Scudder LE, Sullivan CA. Studies with a murine monoclonal antibody that abolishes ristocetin-induced binding of von Willebrand factor to platelets: additional evidence in support of GPIb as a platelet receptor for von Willebrand factor. Blood 1983; 61: 99-110
    PubMedGoogle Scholar
  • 10 Ruan C, Tobelem G, McMichael AJ, Drouet L, Legrand Y, Degos L, Kieffer N, Lee H, Caen JP. Monoclonal antibody to human platelet glycoprotein I. II. Effects on human platelet function. Br J Haematol 1981; 49: 511-519
    CrossrefPubMedGoogle Scholar
  • 11 Fujimoto T, Ohara S, Hawiger J. Thrombin-induced exposure and prostacyclin inhibition of the receptor for factor VIII/von Willebrand factor on human platelets. J Clin Invest 1982; 69: 1212-1222
    CrossrefPubMedGoogle Scholar
  • 12 Ruggeri ZM, Bader R, De Marco L. Glanzmann thrombasthenia: deficient binding of von Willebrand factor to thrombin-stimulated platelets. Proc Natl Acad Sci USA 1982; 79: 6038-6041
    CrossrefPubMedGoogle Scholar
  • 13 Ruggeri ZM, De Marco L, Gatti L, Bader R, Montgomery RR. Platelets have more than one binding site for von Willebrand factor. J Clin Invest 1983; 72: 1-12
    CrossrefPubMedGoogle Scholar
  • 14 Gralnick HR, Williams SB, Coller BS. Fibrinogen competes with von Willebrand factor for binding to the glycoprotein IIb/IIIa complex when platelets are stimulated with thrombin. Blood 1984; 64: 797-800
    PubMedGoogle Scholar
  • 15 Schullek J, Jordan J, Montgomery RR. Interaction of von Willebrand factor with human platelets in the plasma milieu. J Clin Invest 1984; 73: 421-428
    CrossrefPubMedGoogle Scholar
  • 16 Plow EF, Srouji AH, Meyer D, Marguerie G, Ginsberg MH. Evidence that three adhesive proteins interact with a common recognition site on activated platelets. J Biol Chem 1984; 259: 5388-5391
    PubMedGoogle Scholar
  • 17 Stel HV, Sakariassen KS, Scholte BJ, Veerman EC I, van der Kwast TH H, de Groot PG, Sixma JJ, van Mourik JA. Characterization of 25 monoclonal antibodies to factor VIII-von Willebrand factor: relationship between ristocetin-induced platelet aggregation and platelet adherence to subendothelium. Blood 1984; 63: 1408-1415
    PubMedGoogle Scholar
  • 18 Stel HV, Sakariassen KS, de Groot PG, van Mourik JA, Sixma JJ. The von Willebrand factor in the vessel wall mediates platelet adherence. Blood 1985; 65: 85-90
    PubMedGoogle Scholar
  • 19 Meyer D, Baumgartner HR, Edgington TS. Hybridoma antibodies to human von Willebrand factor. II. Relative role of intramolecular loci in mediation of platelet adhesion to subendothelium. Br J Haematol 1984; 57: 609-620
    CrossrefPubMedGoogle Scholar
  • 20 Goodall AH, Jarvis J, Rawlings E, Tuddenham EG D. A specific immunoradiometric assay for VIII: vWF using monoclonal antibodies recognizing a functional site on VIII :RAg. Thromb Haemostas 1983; 50: 111
    PubMedGoogle Scholar
  • 21 Sixma JJ, Sakariassen KS, Stel HV, Houdijk WP M, in der Maur DW, Hamer RJ, de Groot PG, van Mourik JA. Functional domains on von Willebrand factor. Recognition of discrete tryptic fragments by monoclonal antibodies that inhibit interaction of von Willebrand factor with platelets and with collagen. J Clin Invest 1984; 74: 736-744
    CrossrefPubMedGoogle Scholar
  • 22 Nokes TJ C, Mahmoud NA, Savidge GF, Goodall AH, Meyer D, Edgington TS, Hardisty RM. Von Willebrand factor has more than one binding site for platelets. Thromb Res 1984; 34 361: 366
    PubMedGoogle Scholar
  • 23 Houdijk WP M, Schiphorst ME, Sixma JJ. Identification of functional domains on von Willebrand factor by the binding of tryptic fragments to collagen and to platelets in the presence of ristocetin. Blood 1986; 67: 1498-1504
    PubMedGoogle Scholar
  • 24 Fujimura Y, Titani K, Holland LZ, Russel SR, Roberts JR, Elder JH, Ruggeri ZM, Zimmerman TS. Von Willebrand Factor. A reduced and alkylated 52/48 kDa fragment beginning at amino acid residue 449 contains the domain inhereting with platelet glycoprotein Ib. J Biol Chem 1986; 261: 381-385
    PubMedGoogle Scholar
  • 25 Girma JP, Kalfatis M, Pietu G, Lavergne JM, Chopek MW, Edgington TS, Meyer D. Mapping of distinct von Willebrand factor domains interacting with platelet GPIb and GPIIb/IIIa and with collagen using monoclonal antibodies. Blood 1986; 67: 1356-1367
    PubMedGoogle Scholar
  • 26 Bockenstedt P, Greenberg JM, Handin RI. Structural basis of von Willebrand factor binding to glycoprotein Ib and collagen. J Clin Invest 1986; 77: 743-749
    CrossrefPubMedGoogle Scholar
  • 27 van Mourik JA, Mochtar IA. Purification of human anti-hemophilic factor (factor VIII) by gel-chromatography. Biochim Biophys Acta 1970; 221: 677-679
    CrossrefPubMedGoogle Scholar
  • 28 Fraker PJ, Speck JC. Protein and cell membrane iodinations with a sparingly soluble chloramide, 1, 3, 4, 6-tetrachloro-3α, 6α-diphenyl glycoluril. Biochem Biophys Res Commun 1978; 80: 849-857
    CrossrefPubMedGoogle Scholar
  • 29 Sakariassen KS, Bolhuis PA, Sixma JJ. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII- von Willebrand factor bound to the subendothelium. Nature 1979; 279: 635-638
    CrossrefPubMedGoogle Scholar
  • 30 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedGoogle Scholar
  • 31 Phillips DR, Agin DP. Platelet plasma membrane glycoproteins: evidence for the presence of non-equivalent disulfide bonds using nonreduced-reduced two-dimensional gel electrophoresis. J Biol Chem 1977; 252: 2121-2126
    PubMedGoogle Scholar
  • 32 Weiss HJ, Tschopp TB, Baumgartner HR, Sussman II, Johnson MM, Egan JJ. Decreased adhesion of giant (Bernard-Soulier) platelets to subendothelium: further implication on the role of the von Willebrand factor in hemostasis. Am J Med 1974; 57: 920-925
    CrossrefPubMedGoogle Scholar
  • 33 Tschopp TB, Weiss HJ, Baumgartner HR. Interaction of thrombas-thenic platelets with subendothelium: normal adhesion, absent aggregation. Experientia 1975; 31: 113-115
    CrossrefPubMedGoogle Scholar
  • 34 Weiss HJ, Turitto VT, Baumgartner HR. Platelet adhesion and thrombus formation on subendothelium in platelets deficient in glycoproteins IIb-IIIa, Ib and storage granules. Blood 1986; 67: 322-331
    CrossrefPubMedGoogle Scholar
  • 35 Sakariassen KS, Nievelstein PF E M, Coller BS, Sixma JJ. The role of platelet membrane glycoprotein Ib and IIb-IIIa in platelet adherence to human artery subendothelium. Br J Haematol 1986; 63: 681-691
    CrossrefPubMedGoogle Scholar
  • 36 Lopez-Fernandez MF, Ginsberg MH, Ruggeri ZM, Bader FJ, Zimmerman TS. Multimeric structure of platelet factor VIII/von Willebrand factor: the presence of larger multimers and their reassociation with thrombin-stimulated platelet. Blood 1982; 60: 1132-1138
    PubMedGoogle Scholar
  • 37 Turitto VT, Weiss HJ, Baumgartner HR. Platelet interaction with rabbit subendothelium in von Willebrand’s disease: altered thrombus formation distinct from defective platelet adhesion. J Clin Invest 1984; 74: 1730-1741
    CrossrefPubMedGoogle Scholar
  • 38 Titani K, Kumer S, Takio K, Ericsson LM, Wade RD, Ahida K, Walsh KA, Chopek MW, Sadler JE, Fujikawa K. Amino acid sequence of human von Willebrand factor. Biochemistry 1986; 25: 3171-3184
    CrossrefPubMedGoogle Scholar
  • 39 Shelton-Inloes BB, Titani K, Sadler JE. cDNA sequence for human von Willebrand factor reveal five types of repeated domains and five possible protein sequence polymorphins. Biochemistry 1986; 25: 3164-3171
    CrossrefPubMedGoogle Scholar
  • 40 Verwey CL, Diergaarde PJ, Hast M, Pannekoek H. Full length of von Willebrand factor (vWF) cDNA encodes a highly repetitive protein considerably larger than the mature vWF subunit. The EMBO J 1986; 5: 1839-1849
    PubMedGoogle Scholar
  • 41 Pierschbacher MD, Ruoshlati E. Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature 1984; 309: 3033
    PubMedGoogle Scholar
  • 42 Plow EF, Pierschbacher MD, Ruohslahti E, Marguerie GA, Ginsberg MH. The effect of Arg-Gly-Asp containing peptides on fibrinogen and von Willebrand factor to platelets. Proc Natl Acad Sci USA 1985; 82: 8057-8061
    CrossrefPubMedGoogle Scholar
  • 43 Girma JP, Chopek MW, Titani K, Davie EW. Limited proteolysis of human von Willebrand factor by Staphylococcus aureus V8 protease. Isolation and partial characterization of a platelet binding domain. Biochemistry 1986; 25: 3156-3163
    CrossrefPubMedGoogle Scholar