Ristocetin and Botrocetin Involve Two Distinct Domains of von Willebrand Factor for Binding to Platelet Membrane Glycoprotein lb

Summary

We have evidence that ristocetin and botrocetin mediate binding of von Willebrand Factor (vWF) to platelet glycoprotein lb (GPIb) through two distinct domains on the vWF molecule. This was established by using monoclonal antibodies (MAbs) to vWF and synthetic peptides derived from the sequence of vWF. MAb 322 and MAb NMC/vW 4 both recognize native vWF as well as fragments containing the GPIb-binding domain of vWF, obtained with the following enzymes: trypsin (116 kDa), V-8 pro tease (Spill, 320 kDa) and V-8 protease plus subtilisin (33-28 kDa). Nevertheless, the lack of reciprocal displacement between the two MAbs in experiments of competitive inhibition for binding to vWF demonstrate that their respective epitopes are separate. Both MAbs inhibit ¹²⁵I-vWF binding to platelet membrane GPIb and vWF-dependent platelet agglutination induced by ristocetin. However, only MAb NMC/vW4 inhibits these functions in the presence of botrocetin and when ristocetin-induced platelet agglutination is inhibited by MAb 322, botrocetin is still able to restore the agglutination. The involvement of two distinct domains of vWF for binding to GPIb in the presence of ristocetin or botrocetin was confirmed in experiments of binding of ¹²⁵I-vWF to platelets using as competitor synthetic peptides corresponding to the GPIb binding domain of vWF (Cys 474 to Pro 488 and Ser 692 to Pro 708). At a final concentration of 2.5 mM both peptides inhibit more than 90% of the binding of vWF to ristocetin-treated platelets but are unable to modify this binding in the presence of botrocetin. In conclusion our data suggest that botrocetin and ristocetin involve distinct sites on vWF for binding to GPIb.

• References

• 1 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 Bemard-Soulier syndrome. J Lab Clin Med 1978; 92: 750-764
• 2 Baumgartner HR, Tschopp TB, Meyer D. Shear rate dependent inhibition of platelet adhesion and agglutination on collagenous surfaces by antibodies to human factor VUI/von Willebrand factor. Br J Haematol 1980; 44: 127-139
• 3 Kao KJ, Pizzo SV, McKee PA. Demonstration and characterization of specific binding sites for factor VUI/von Willebrand factor on human platelets. J Clin Invest 1979; 63: 656-664
• 4 Moake JL, Olson JD, Troll JH, Tang SS, Funicella T, Peterson DM. Binding of radioiodinated human von Willebrand factor to Bemard-Soulier, thrombasthenic and von Willebrand’s disease platelets. Thromb Res 1980; 19: 21-27
• 5 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
• 6 Ruan C, Tobelem G, McMichael AJ, Drouet L, Legrand Y, Degos L, Kieffer N, Lee H, Caen JP. Monoclonal antibody to human platelet glycoprotein 1. II Effects on human platelet function. Br J Haematol 1981; 49: 511-519
• 7 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
• 8 Read MS, Shermer RW, Brinkhous KM. Venom coagglutinin: an activator of platelet aggregation dependent on von Willebrand factor. Proc Natl Acad Sci USA 1978; 75: 4514-4518
• 9 Brinkhous KM, Read MS. Use of venom coagglutinin and lyophilized platelets in testing for platelet-aggregating von Willebrand factor. Blood 1980; 55: 517-520
• 10 Chopek MW, Girma JP, Fujikawa K, Davie EW, Titani K. Human von Willebrand factor: a multivalent protein composed of identical subunits. Biochemistry USA 1986; 25: 3146-3155
• 11 Titani K, Kumar S, Takio K, Ericsson LH, Wade RD, Ashida K, Walsh KA, Chopek MW, Sadler JE, Fujikawa K. Amino acid sequence of human von Willebrand factor. Biochemistry USA 1986; 25: 3171-3184
• 12 Sixma JJ, Sakariassen KS, Stel HV, Houdijk W PM, in der Maur DW, Hamer RJ, deGroot PG, van Mourik JA. Functional domains of von Willebrand factor. Recognition of discrete tryptic fragments by monoclonalantibodies that inhibit interaction of von Willebrand factor with platelets and with collagen. J Clin Invest 1984; 74: 736-744
• 13 Girma JP, Chopek MW, Titani K, Davie EW. Limited proteolysis of human von Willebrand factor by staphylococcus aureus V-8 protease: isolation and partial characterization of a platelet-binding domain. Biochemistry USA 1986; 25: 3156-3163
• 14 Girma JP, Kalafatis 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: 1356-1366
• 15 Fujimura Y, Titani K, Holland LZ, Russell 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 interacting with platelet glycoprotein Ib. J Biol Chem 1986; 261: 381-385
• 16 Fujimura Y, Holland LZ, Ruggeri ZM, Zimmerman TS. The von Willebrand factor domain-mediating botrocetin-induced binding to glycoprotein lb lies between Val 449 and Lys 728. Blood 1986; 70: 985-988
• 17 Mohri H, Fujimura Y, Shima M, Yoshioka A, Houghten RA, Ruggeri ZM, Zimmerman TS. Structure of the von Willebrand factor domain interacting with glycoprotein Ib. J Biol Chem 1988; 263: 17901-17904
• 18 Howard MA, Perkin J, Salem HH, Firkin BG. The agglutination of human platelets by botrocetin: evidence that botrocetin and ristocetin act at different sites on the factor VIII molecule and platelet membrane. Br J Haematol 1984; 57: 25-35
• 19 Thorell L, Blomback B. Purification of the factor VIII complex. Thromb Res 1984; 35: 431-50
• 20 Takahashi Y, Kalafatis M, Girma JP, Sewerin K, Andersson LO, Meyer D. Localization of a factor VIII binding domain on a 34 kilodalton fragment of the N-terminal portion of von Willebrand factor. Blood 1987; 70: 1679-1682
• 21 Shima M, Morimoto J, Imai S, Tsubura Y, Yoshioka A, Fukui H. Production and characterization of monoclonal antibodies against von Willebrand factor (vWF). J Nara Med Assoc 1985; 36: 662-669
• 22 Fine JM, Steinbuch M. A simple technique forthe isolation of monoclonal IgG and IgA. Rev Eur Et Clin Biol 1970; 15: 1115-1121
• 23 Kalafatis M, Takahashi Y, Girma JP, Meyer D. Localization of a collagen-interactive domain of human von Willebrand factor between amino acid residues Gly 911 and Glu 1,365. Blood 1987; 70: 1577-1583
• 24 Merrifield RB, Steward JM, Jernberg N. Instrument for automated synthesis of peptides. Anal Chem 1966; 38: 1905-1914
• 25 Fraker PJ, Speck JC. Proteins and cell membrane iodinations with a sparingly soluble chloroamide, 1, 3, 4, 6-tetrachloro-3a, 6a diphenyl-glycoluril. Biochem Biophys Res Comm 1978; 80: 849-857
• 26 Pietu G, Fressinaud E, Girma JP, Nieuwenhuis HK, Rothschild C, Meyer D. Binding of von Willebrand factor to collagen and to collagen-stimulated platelets. J Lab Clin Med 1987; 109: 637-646
• 27 Allain JP, Cooper HA, Wagner RH, Brinkhous KM. Platelets fixed with paraformaldehyde: a new reagent for assay of von Willebrand factor and platelet aggregating factor. J Lab Clin Med 1975; 85: 318-328
• 28 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970; 227: 680-685
• 29 Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of protein from polyacrylamide gels to nitrocellulose sheets. Procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
• 30 De Groot PG, Ottenhof-Rovers M, van Mourik JA, Sixma JJ. Evidence that the primary binding site of von Willebrand factor that mediates platelet adhesion on subendothelium is not collagen. J Clin Invest 1988; 82: 65-73
• 31 Sakariassen KS , Bolhuis P, Sixma JJ. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII - von Willebrand factor bound to the subendothelium. Nature 1979; 279: 636-638
• 32 Andrews RK, Gorman JJ, Booth WJ, Corino GL, Castaldi PA, Berndt MC. Cross-linking of a monomeric 39/34 kDa dispase fragment of von Willebrand factor (Leu-480/Val 481-Gly-718) to the N-terminal region of the a-chain of membrane glycoprotein I b on intact platelets with bis(sulfosuccinimidyl) suberate. Biochemistry USA 1989; 28: 8326-8336
• 33 Read MS, Smith SV, Lamb MA, Brinkhous KM. Role of botrocetin in platelet agglutination: formation of an activated complex of botrocetin and von Willebrand factor. Blood 1989; 74: 1031-1035
• 34 Nishio K, Fujimura Y, Mizumoto Y, Fukui H, Tomiyama Y, Kurata Y, Tsubakio T, Yonezawa Y, Usami Y, Titani K. A new anti-platelet glycoprotein (GP)Ib monoclonal antibody (OP-F1) demonstrates the non-identical von Willebrand Factor binding sites on the GPIb molecule expressed by ristocetin or botrocetin. Thromb Haemostas 1989 62. 219 (Abstr)
• 35 Brinkhous KM, Read M S, Fricke WA, Wagner RH. Botrocetin (venom coagglutinin): reaction with a broad spectrum of multimeric forms of factor VIII macromolecular complex. Proc Natl Acad Sci USA 1983; 80: 1463-1466
• 36 Andrews RK, Booth WJ, Gorman JJ, Castaldi PA, Berndt MC. Purification of botrocetin from Bothrops jararaca venom. Analysis of the botrocetin-mediated interaction between von Willebrand Factor and the human platelet membrane glycoprotein Ib-IX complex. Biochemistry USA 1989; 28: 8317-8326