Download PDF
Thromb Haemost 1997; 77(01): 174-182
DOI: 10.1055/s-0038-1655927
von Willebrand Factor
Schattauer GmbH Stuttgart

Abnormal Proteolytic Processing of von Willebrand Factor Arg611 Cys and Arg 611 His

Toshiya Nishikubo
1   The Inserm U.143, Hôpital de Bicêtre, Paris, France
,
Olivier Christophe
1   The Inserm U.143, Hôpital de Bicêtre, Paris, France
,
Jean-Maurice Lavergne
1   The Inserm U.143, Hôpital de Bicêtre, Paris, France
,
Bernadette Obert
1   The Inserm U.143, Hôpital de Bicêtre, Paris, France
,
Kyoko Nonami
2   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Yukihiro Takahashi
2   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Akira Yoshioka
2   Department of Pediatrics, Nara Medical University, Kashihara, Nara, Japan
,
Dominique Meyer
1   The Inserm U.143, Hôpital de Bicêtre, Paris, France
,
Jean-Pierre Girma
1   The Inserm U.143, Hôpital de Bicêtre, Paris, France
› Author Affiliations
Further Information

Publication History

Received 08 May 1996

Accepted after revision 30 September 1996

Publication Date:
11 July 2018 (online)

Also available at
  PDF Download  Permissions and Reprints

Summary

The structural and functional properties of plasma and platelet vWF were studied in 8 patients (5 unrelated families) with vWD demonstrating a mutation at position 611 (R611C or R611H). Following reduction, electrophoresis and immunoblotting with a polyclonal anti-reduced vWF antibody, abnormal proteolysis of vWF was demonstrated in plasma and to a lesser extent in platelets from all patients, leading to the formation of a unique 209 kDa fragment undetectable in control as well as in type 2A, 2B or 2N vWF. Immunoblotting with MoAbs to reduced vWF showed that the C-terminal end of the 209 kDa fragment was located beyond residue 1744 of the subunit and that its N-terminus was between residues 523 and 1114. Multimeric analysis of patients vWF showed an abnormal pattern in both plasma and platelets, with a moderate decrease of the HMW multimers together with a significant increase of the lowest MW forms. The specific sensitivity of vWF R611C and vWF R611H to proteolysis was further evidenced using V-8 protease. In all patient’s samples the enzyme produced a unique monomeric 80 kDa fragment, absent in V-8 digested normal vWF, which overlapped the N-terminal part of the subunit. The functional analysis of vWF showed a markedly decreased affinity of mutated plasma vWF for platelet GPIb in the presence of ristocetin. Infusion of DDAVP in two of these patients did not lead to significant platelet count change. It induced a limited increase of the HMW multimers in plasma together with a poor correction of the vWF binding to platelet GPIb. In conclusion, our data demonstrate that in addition to a normal proteolysis, vWF mutated at position 611 undergoes a specific cleavage in plasma and platelets. In contrast to the increased proteolysis observed in type 2A and 2B patients’ plasma, this additional cleavage produced a unique 209 kDa species but maintained a HMW multimer-like structure of vWF R611C and R611H.

 

  • References

  • 1 Ruggeri ZM, Ware J. The structure and function of von Willebrand Factor. Thromb Haemost 1992; 67: 594-599
    Thieme ConnectPubMedSearch in Google Scholar
  • 2 Meyer D, Girma JP. von Willebrand Factor – structure and function. Thromb Haemost 1993; 70: 99-104
    Thieme ConnectPubMedSearch in Google Scholar
  • 3 Chopek MW, Girma JP, Fujikawa K, Davie EW, Titani K. Human von Willebrand Factor: a multivalent protein composed of identical subunits. Biochemistry 1986; 25: 3146-3155
    CrossrefPubMedSearch in Google Scholar
  • 4 Titani K, Kumar S, Ericsson LH, Wade RD, Ashida K, Walsh KA, Chopek MW, Sadler JE, Fujikawa K. Amino acid sequence of human von Willebrand Factor. Biochemistry 1986; 25: 3171-3184
    CrossrefPubMedSearch in Google Scholar
  • 5 Andrews RK, Gorman JJ, Booth WJ, Corino GL, Castaldi PA, Bemdt MC. Cross linking of a monomeric 39/34 kDa dispase fragment of von Willebrand Factor (Leu480/Val481 -Gly718) to the N-terminal region of the œ-chain of membrane glycoprotein Ib on the intact platelets with bis (sulfosuccinimidyl) suberate. Biochemistry 1989; 28: 8326-8336
    CrossrefPubMedSearch in Google Scholar
  • 6 Marti T, Rösselet SJ, Titani K, Walsh KA. Identification of disulfide- bridged substructures within human von Willebrand Factor. Biochemistry 1987; 26: 8099-8109
    CrossrefPubMedSearch in Google Scholar
  • 7 Zimmerman TS, Dent JA, Ruggeri ZM, Nannini LH. Subunit composition of plasma von Willebrand Factor. Cleavage is present in normal individuals, increased in type IIA and IIB von Willebrand disease, but minimal in variants with aberrant structure of individual oligomers (types IIC, IID, and IIE). J Clin Invest 1986; 77: 947-951
    CrossrefPubMedSearch in Google Scholar
  • 8 Dent JA, Galbusera M, Ruggeri ZM. Heterogeneity of plasma von Willebrand Factor multimers resulting from proteolysis of the constituent subunit. J Clin Invest 1991; 88: 774-782
    CrossrefPubMedSearch in Google Scholar
  • 9 Furlan M, Robles R, Lammle B. Partial purification and characterization of a protease from human plasma cleaving von Willebrand Factor to fragments produced by in vivo proteolysis. Blood 1996; 87: 4223-4234
    CrossrefPubMedSearch in Google Scholar
  • 10 Tsai HM. Physiologic cleavage of von Willebrand Factor by a plasma protease is dependent on its conformation and requires calcium ion. Blood 1996; 87: 4235-4244
    PubMedSearch in Google Scholar
  • 11 Berkowitz SD, Dent J, Roberts J, Fujimura Y, Plow EF, Titani K, Ruggeri ZM, Zimmerman TS. Epitope mapping of the von Willebrand Factor subunit distinguishes fragments present in normal and type IIA von Willebrand disease from those generated by plasmin. J Clin Invest 1987; 79: 524-531
    CrossrefPubMedSearch in Google Scholar
  • 12 Dent JA, Berkowitz SD, Ware J, Kasper CK, Ruggeri ZM. Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand Factor. Proc Natl Acad Sci USA 1990; 87: 6306-63010
    CrossrefPubMedSearch in Google Scholar
  • 13 Sadler JE. A revised classification of von Willebrand disease. Thromb Haemost 1994; 71: 520-525
    Thieme ConnectPubMedSearch in Google Scholar
  • 14 Lyons SE, Bruck ME, Bowie EJW, Ginsburg D. Impaired intracellular transport produced by a subset of type-IIA von Willebrand disease mutations. J Biol Chem 1992; 267: 4424-4430
    PubMedSearch in Google Scholar
  • 15 Hakami N, Shibuya H, Johnson GS, Stoy SJ, Huang THM, Montgomery RR. A novel von Willebrand Factor, Arg611Cys or vWF Columbia, associated with a dominant von Willebrand disease variant. Blood 1993; 82: 149a (abstr)
    PubMedSearch in Google Scholar
  • 16 Castaman G, Eikenboom JCJ, Rodeghiero F, Briët E, Reitsma PH. A novel candidate mutation (Arg(611)→His) in type I ‘platelet discordant’ von Willebrand’s disease with desmopressin-induced thrombocytopenia. Br J Haematol 1995; 89: 656-658
    CrossrefPubMedSearch in Google Scholar
  • 17 Hilbert L, Gaucher C, Mazurier C. Identification of two mutations (Arg611Cys and Arg61 lHis) in the A1 loop of von Willebrand Factor (vWF) responsible for type 2 von Willebrand disease with decreased platelet-dependent function of vWF. Blood 1995; 86: 1010-1018
    PubMedSearch in Google Scholar
  • 18 Weiss HJ, Pietu G, Rabinowitz R, Girma JP, Rogers J, Meyer D. Heterogeneous abnormalities in the multimeric structure, antigenic properties, and plasma-platelet content of factor VIII/von Willebrand Factor in subtypes of classic (type I) and variant (type IIA) von Willebrand’s disease. J Lab Clin Med 1983; 101: 411-425
    PubMedSearch in Google Scholar
  • 19 Nishino M, Girma JP, Rothschild C, Fressinaud E, Meyer D. New variant of von Willebrand disease with defective binding to Factor VIII. Blood 1989; 05: 1591-1599
    PubMedSearch in Google Scholar
  • 20 Weiss HJ, Meyer D, Rabinowitz R, Pietu G, Girma JP, Vicic WJ, Rogers J. Pseudo-von Willebrand’s disease. An intrinsic platelet defect with aggregation by unmodified human Factor VIII/von Willebrand Factor and enhanced adsorption of its high-molecular-weight multimers. N Engl J Med 1982; 306: 326-333
    CrossrefPubMedSearch in Google Scholar
  • 21 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
    PubMedSearch in Google Scholar
  • 22 Meyer D, Zimmerman TS, Obert B, Edgington TS. Hybridoma antibodies to human von Willebrand Factor.I. Characterization of seven clone. Br J Haematol 1984; 57: 597-608
    CrossrefPubMedSearch in Google Scholar
  • 23 Christophe O, Ribba AS, Baruch D, Obert B, Rouault C, Niinomi K, Pietu G, Meyer D, Girma JP. Influence of mutations and size of multimers in type II von Willebrand disease upon the function of von Willebrand Factor. Blood 1994; 83: 3553-3561
    PubMedSearch in Google Scholar
  • 24 Pietu G, Ribba A S, Cherel G, Siguret V, Obert B, Rouault C, Ginsburg D, Meyer D. Epitope mapping of inhibitory monoclonal antibodies to human von Willebrand Factor by using recombinant cDNA libraries. Thromb Haemost 1994; 71: 788-792
    Thieme ConnectPubMedSearch in Google Scholar
  • 25 Ardaillou N, Girma JP, Meyer D, Lavergne JM, Shoa’i I, Larrieu MJ. “Variants” of von Willebrand’s disease. Demonstration of a decreased antigenic reactivity by immunoradiometric assay. Thromb Res 1978; 12: 817-830
    CrossrefPubMedSearch in Google Scholar
  • 26 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
    CrossrefPubMedSearch in Google Scholar
  • 27 Christophe O, Obert B, Meyer D, Girma JP. The binding domain of von Willebrand Factor to sulfatides is distinct from those interacting with glycoprotein Ib heparin, and collagen and resides between amino acid residues Leu512 and Lys673. Blood 1991; 78: 2310-2317
    PubMedSearch in Google Scholar
  • 28 Christophe O, Rouault C, Obert B, Pietu G, Meyer D, Girma JP. A monoclonal antibody (B724) to von Willebrand Factor recognizing an epitope within the A1 disulphide loop (Cys509-Cys695) discriminates between type 2A and type 2B von Willebrand disease. Br J Haematol 1995; 90: 195-203
    CrossrefPubMedSearch in Google Scholar
  • 29 Takahashi Y, Kalafatis M, Girma JP, Meyer D. Abnormality of the N-terminal portion of von Willebrand Factor in type IIA and IIC von Willebrand disease. Thromb Haemost 1988; 60: 498-505
    Thieme ConnectPubMedSearch in Google Scholar
  • 30 Laemmli UK. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 1970; 227: 680-685
    CrossrefPubMedSearch in Google Scholar
  • 31 O’Farrel PH. High resolution two-dimensional electrophoresis of proteins. J Biol Chem 1975; 250: 4007-4021
    PubMedSearch in Google Scholar
  • 32 Towbin H, Stachelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci USA 1979; 76: 4350-4354
    CrossrefPubMedSearch in Google Scholar
  • 33 Johnson DA, Gautsch JW, Sportman JR, Elder JH. Improved technique utilizing nonfat dry milk for analysis of proteins and nucleic acids transferred to nitrocellulose. Gene Anal Techn 1984; 01: 3-8
    CrossrefPubMedSearch in Google Scholar
  • 34 Hancock K, Tsang VCW. India ink staining of proteins on nitrocellulose paper. Anal Biochem 1983; 133: 157-162
    CrossrefPubMedSearch in Google Scholar
  • 35 Ruggeri ZM, Zimmerman TS. The complex multimeric composition of Factor VUI/von Willebrand Factor. Blood 1981; 57: 1140-1143
    PubMedSearch in Google Scholar
  • 36 Fraker PJ, Speck JC. Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a, 6a-diphenylglycoluril. Biochem Biophys Res Commun 1978; 80: 849-857
    CrossrefPubMedSearch in Google Scholar
  • 37 Ribba AS, Lavergne JM, Bahnak BR, Derlon A, Pietu G, Meyer D. Duplication of a methionine within the glycoprotein Ib binding domain of von Willebrand Factor detected by denaturing gradient gel electrophoresis in a patient with type-IIB von Willebrand disease. Blood 1991; 78: 1738-1743
    PubMedSearch in Google Scholar
  • 38 Lavergne JM, Piao Y, Ribba AS, Girma JP, Siguret V, Pietu G, Boyer-Neumann C, Schandelong A, Bahnak BR, Meyer D. Functional analysis of the Arg91Gln substitution in the Factor-VIII binding domain of von Willebrand Factor demonstrates variable phenotypic expression. Thromb Haemost 1993; 70: 691-696
    Thieme ConnectPubMedSearch in Google Scholar
  • 39 Sambrook J, Friitsch EF, Maniatis T. Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Publications. Cold Spring Harbor. NY: 1989
    Search in Google Scholar
  • 40 Batlle J, Lasierra J, Villamor AF, Navarro JL, Pardo A, Campos M, Justica B, Fernandez MFL. Proteolytic processing of von Willebrand Factor subunit – heterogeneity in Type-IIA von Willebrand disease. Ann Hematol 1994; 68: 111-115
    CrossrefPubMedSearch in Google Scholar
  • 41 Levene RB, Booyse FM, Chediak J, Zimmerman TS, Livingston DM, Lynch DC. Expression of abnormal von Willebrand Factor by endothelial cells from a patient with type IIA von Willebrand disease. Proc Natl Acad Sci USA 1987; 84: 6550-6554
    CrossrefPubMedSearch in Google Scholar
  • 42 Foster PA, Fulcher CA, Marti T, Titani K, Zimmerman TS. A major Factor VIII binding domain resides within the amino-terminal 272 amino acid residues of von Willebrand Factor. J Biol Chem 1987; 262: 8443-8446
    CrossrefPubMedSearch in Google Scholar