Primary Binding Domain of Bovine von Willebrand Factor Fragment Expressed in E. coli

Summary

Bovine vWF cDNA has been cloned from a bovine endothelial cell library. A fragment of this cDNA, corresponding to amino acid sequence Leu 469-Ser 723, called primary adhesion domain (PAD-1), and containing the binding sites for platelet glycoprotein Ib (GPIb), heparin and collagen, has been expressed in E. coli. The reduced and alkylated form of fragment PAD-1 inhibited native vWF binding to GPIb. Fragment PAD-1 bound to heparin and botrocetin in a specific and dose dependent manner as did the native vWF. In a solid-phase assay, fragment PAD-1 bound to calf skin collagen in contrast to a human vWF recombinant fragment (Ser 445-Val 733) which was inactive in the same assay. The studies presented in this paper demonstrated that the A1 domain of bovine vWF contained the GPIb, heparin, botrocetin as well as collagen binding sites and that integrity of the disulfide bond (Cys 509-Cys 695), did not seem to be essential for binding of bovine vWF fragment to GPIb.

• References

• 1 Jaffe EA, Hoyer LW, Nachman RL. Synthesis of antihemophilic factor antigen by cultured human endothelial cells. J Clin Invest 1973; 52: 2757-2764
• 2 Nachman R, Levine R, Jaffe EA. Synthesis of Factor VIII antigen by cultured guinea pig megakaryocytes. J Clin Invest 1977; 60: 914-921
• 3 Weiss HJ, Sussman II, Hoyer L. Stabilization of factor VIII in plasma by the von Willebrand factor: Studies on post-transfusion and dissociated factor VIII and in patients with von Willebrand’s disease. J Clin Invest 1977; 60: 390-404
• 4 Sakariassen KS, Bolhuis PA, Sixma JJ. Human blood platelet adhesion to artery subendothelium is mediated by factor VIII-vWF bound to the subendothelium. Nature 1979; 279: 636-638
• 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: 01-12
• 6 Ruggeri ZM, Ware J. von Willebrand factor. [Review] FASEB Journal 1993; 7: 308-316
• 7 Meyer D, Girma JP. von Willebrand factor: structure and function. Thromb Haemost 1993; 70: 099-104
• 8 Sadler EJ. von Willebrand factor, minireview. J Biol Chem 1991; 266: 22777-22780
• 9 Howard MA, Firkin BG. Ristocetin - a new tool in the investigation of platelet aggregation. Thromb Haemost 1971; 26: 362-369
• 10 Kao KJ, Pizzo SV, McKee PA. Demonstration and characterization of specific binding sites for factor VIII/ von Willebrand factor on human platelets. J Clin Invest 1979; 63: 656-664
• 11 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
• 12 Andrews RK, Booth WJ, Gorman JJ, Castaldi PA, Bemdt 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 1989; 28: 8317-8325
• 13 Read MS, Smith SV, Lamb MA, Brinkhous KM. Role of botroctin in platelet agglutination: Formation of an activated complex of botrocetin and von Willebrand factor. Blood 1989; 74: 1031-1035
• 14 Mohri H, Yoshioka A, Zimmerman TS, Ruggeri ZM. Isolation of the von Willebrand factor domain interacting with platelet glycoprotein Ib, heparin, and collagen, and characterization of its three distinct functional sites. J Biol Chem 1989; 264: 17361-17367
• 15 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 1986; 25: 3156-3163
• 16 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 interacting with platelet glycoprotein Ib. J Biol Chem 1986; 261: 381-385
• 17 Sugimoto M, Ricca G, Hrinda ME, Schreiber AB, Searfoss G, Bottini E, Ruggeri ZM. Functional modulation of the isolated glycoprotein lb binding domain of von Willebrand factor expressed in Escherichia coli . Biochemistry 1991; 30: 5202-5209
• 18 Sugimoto M, Dent J, McClintock R, Ware J, Ruggeri ZM. Analysis of structure-function relationships in the platelet membrane glycoprotein Ib-binding domain of von Willebrand’s factor by expression of deletion mutants. J Biol Chem 1993; 268: 12185-12192
• 19 Pietu G, Muellen P, Cherel G, Diaz J, Baruch D, Cortney M, Meyer D. Production in Escherichia coli of a biologically active subfragment of von Willebrand factor corresponding to the platelet glycoprotein Ib, collagen and heparin domains. Biochem Biophys Res Comm 1989; 164: 1339-1347
• 20 Prior C, Chu V, Holt J, Windisch V, Lee T, Mitschelen J, Newman J, Ricca G, Tarr C, Hrinda ME. Production and functional characterization of a recombinant fragment of von Willebrand Factor (vWF): An antagonist to platelet receptor GP Ib. Biotechnology 1992; 10: 66-73
• 21 Gralnick HR, Williams S, McKeown L, Kramer W, Krutzsch H, Górecki M, Pinet A, Garfinkel LI. A monomeric von Willebrand factor fragment, Leu 504-Ser 728, inhibits von Willebrand factor interaction with glycoprotein Ib-IX. Proc Natl Acad Sci 1992; 89: 7880-7884
• 22 Cruz MA, Handin RI, Wise RJ. The interaction of the von Willebrand fac-tor-Al domain with platelet glycoprotein MX. The role of glycosylation and disulfide bonding in a monomeric recombinant A1 domain protein. J Biol Chem 1993; 268: 21238-21245
• 23 Azuma H, Dent JA, Sugimoto M, Ruggeri ZM, Ware J. Independent assembly and secretion of a dimeric adhesive domain of von Willebrand factor containing the glycoprotein Ib-binding site. J Biol Chem 1991; 266: 12342-12347
• 24 Bemdt MC, Ward CM, Boorth WJ, Castaldi PA, Mazurov AV, Andrews RK. Identification of aspartic acid 514 through glutamic acid 542 as a glycoprotein Ib-IX complex receptor recognition sequence in von Willebrand factor. Mechanism of modulation of von Willebrand factor by ristocetin and botrocetin. Biochemistry 1992; 31: 11144-11151
• 25 Kirby EP. The agglutination of human platelets by bovine factor VIII:R. J Lab Clin Med 1982; 100: 963-976
• 26 Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1960; 227: 680-685
• 27 Fairbanks G, Steck TL, Wallach DFH. Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 1971; 10: 2606-2617
• 28 Bakhshi MR, Myers JC, Howard PS, Soprano DR, Kirby EP. Sequencing of the primary adhesion domain of bovine von Willebrand factor. Biochim Biophys Acta 1992; 1132: 325-328
• 29 Mascelli MA, Edgington TS, Kirby EP. Characterization of a fragment of bovine von Willebrand factor that binds to platelets. Biochemistry 1986; 25: 6325-6335
• 30 Fraker PJ, Speck JC. Protein and cell membrane iodinations with sparingly soluble chloramid, 1,3,4,6-tetrachloro-3α, 6α-diphenylglycouril. Biochem Biophys Res Commun 1978; 80: 849-857
• 31 Smith JW, Knauer DJ. Ligand blotting with ¹²⁵I fluoresceinamine-heparin. Anal Biochem 1987; 160: 105-114
• 32 Mustard JF, Perry DW, Ardlie NG, Packham MA. Preparation of suspensions of washed platelets of humans. Br J Haematol 1972; 22: 193-204
• 33 Kirby EP, Mills DCB. The interaction of bovine factor VIII with human platelets. J Clin Invest 1975; 56: 591-602
• 34 Bakhshi MR, Kirby EP. Proteolytic and immunologic comparison of human and bovine von Willebrand factor. Thromb Haemost 1990; 65: 517-523
• 35 Girma JP, Ribba AS, Meyer D. Structure-function relationship of the A1 domain of von Willebrand factor. Thromb Haemost 1995; 74: 156-160
• 36 Sixma JJ, Schiphorst ME, Verweij CL, Pannekoek H. Effect of deletion of the A1 domain of von Willebrand factor on its binding to heparin, collagen and platelets in the presence of ristocetin. Eur J Biochem 1991; 196: 369-375
• 37 Sugimoto M, Mohri H, McClintock RA, Ruggeri ZM. Identification of discontinuous von Willebrand factor sequences involved in complex formation with botrocetin. J Biol Chem 1991; 266: 18172-18178
• 38 De Marco L, Girolami A, Russell S, Ruggeri ZM. Interaction of asilo von Willebrand factor with glycoprotein Ib induces fibrinogen binding to the glycoprotein IMIIa complex and mediates platelet aggregation. J Clin Invest 1985; 75: 1198-1203
• 39 Knott HM, Bemdt MC, Kralicek AV, O’Donoghue SI, King GF. Determination of the solution structure of a platelet-adhesion peptide of von Willebrand factor. Biochemistry 1992; 31: 11152-11158
• 40 Carew JA, Quinn SM, Stoddart JH, Lynch DC. O-linked carbohydrate or recombinant von Willebrand factor influences ristocetin-induced binding to platelet glycoprotein Ib. J Clin Invest 1992; 90: 2258-2267