Download PDF
Thromb Haemost 2013; 109(06): 999-1006
DOI: 10.1160/TH12-11-0808
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

C2362F mutation gives rise to an ADAMTS13-resistant von Willebrand factor

Alessandra Casonato
1   University of Padua Medical School, Department of Cardiologic, Thoracic and Vascular Sciences, Second Chair of Internal Medicine, Padua, Italy
,
Elena Pontara
1   University of Padua Medical School, Department of Cardiologic, Thoracic and Vascular Sciences, Second Chair of Internal Medicine, Padua, Italy
,
Monica Battiston
2   Department of Laboratory Diagnostics and Cellular Therapy, IRCCS, C.R.O., Aviano, Pordenone, Italy
,
Margherita Morpurgo
3   Pharmaceutical Chemistry and Pharmacology Department, University of Padua, Italy
,
Maria Grazia Cattini
1   University of Padua Medical School, Department of Cardiologic, Thoracic and Vascular Sciences, Second Chair of Internal Medicine, Padua, Italy
,
Elisabetta Casarin
3   Pharmaceutical Chemistry and Pharmacology Department, University of Padua, Italy
,
Giorgia Saga
1   University of Padua Medical School, Department of Cardiologic, Thoracic and Vascular Sciences, Second Chair of Internal Medicine, Padua, Italy
,
Viviana Daidone
1   University of Padua Medical School, Department of Cardiologic, Thoracic and Vascular Sciences, Second Chair of Internal Medicine, Padua, Italy
,
Luigi De Marco
2   Department of Laboratory Diagnostics and Cellular Therapy, IRCCS, C.R.O., Aviano, Pordenone, Italy
› Author Affiliations
Further Information

Publication History

Received: 08 November 2012

Accepted after major revision: 05 February 2013

Publication Date:
22 November 2017 (online)

Also available at
    Permissions and Reprints

Summary

von Willebrand factor (VWF) multimers result from proteolysis by the metalloprotease ADAMTS13. Since C2362F-VWF features abnormally large multimers with their triplet oligomer structure replaced by a diffuse smear, we explored the susceptibility of C2362F-VWF to ADAMTS13. VWF-enriched blood samples, obtained by cryoethanol precipitation of plasma from a patient with von Willebrand disease (VWD) homozygous for the C2362F mutation and a normal subject, were submitted to cleavage by recombinant ADAMTS13 under static conditions in the presence of urea. C2362F-VWF proved completely ADAMTS13-resistant in vitro. At any concentration of recombinant ADAMTS13 (from 0.1 μM to 1 μM), there was no evidence of the abnormally large VWF multimers of C2362F-VWF disappearing, nor any increased representation of triplet multimer bands, unlike the situation seen in normal VWF. This is due partly to a defective ADAMTS13 binding to C2362F-VWF under static conditions, as seen in both the patient’s and recombinant mutated VWF proteins. These findings were associated with a significantly shorter than normal survival of C2362F-VWF after DDAVP, demonstrating that proteolysis and VWF survival may be independent phenomena. Our findings clearly demonstrate that the loss of cysteine 2362 makes VWF resistant to proteolysis by ADAMTS13, at least partly due to an impaired ADAMTS13 binding to VWF. This suggests that the B2 domain of VWF is involved in modulating ADAMTS13 binding to VWF and the consequent proteolytic process. The C2362F-VWF mutation also enables a new abnormality to be identified in the VWF-ADAMTS13 relationship, i.e. an ADAMTS13-resistant VWF.

Keywords

VWF - C2362F VWF mutation - VWD - ADAMTS13 - ADAMTS13 binding to VWF
 

  • References

  • 1 Sadler JE. Biochemistry and genetics of von Willebrand factor. Annu Rev Biochem 1998; 67: 395-424.
    CrossrefPubMedGoogle Scholar
  • 2 Ruggeri ZM. Structure and function of von Willebrand factor. Thromb Haemost 1999; 82: 576-584.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 3 Wagner DD, Marder VJ. Biosynthesis of von Willebrand factor protein by human endothelial cells: processing steps and their intracellular localization. J Cell Biol 1984; 99: 2123-2130.
    CrossrefPubMedGoogle Scholar
  • 4 Gralnick HR, Williams SB, Morisato DK. Effect of multimeric structure of the factor VIII/von Willebrand factor protein on binding to platelets. Blood 1981; 58: 387-392.
    PubMedGoogle Scholar
  • 5 Furlan M. Von Willebrand factor: molecular size and functional activity. Ann Hematol 1996; 72: 341-348.
    CrossrefPubMedGoogle Scholar
  • 6 Dent JA, Berkowitz SD, Ware J. et al. 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: 6306-6310.
    CrossrefPubMedGoogle Scholar
  • 7 Furlan M, Robles R, Lamie B. Partial purification and characterization of a protease from human plasma cleaving von Willebrand factor to fragments produced by an in vivo proteolysis. Blood 1996; 87: 4223-4234.
    PubMedGoogle Scholar
  • 8 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.
    PubMedGoogle Scholar
  • 9 Zheng X, Chung D, Takayama TK. et al. Structure of von Willebrand factor-cleaving protease (ADAMTS13), a metalloprotease involved in thrombotic thrombocytopenic purpura. J Biol Chem 2001; 276: 41059-41063.
    CrossrefPubMedGoogle Scholar
  • 10 Crawley JT, de Groot R, Xiang Y. et al. Unravelling the scissile bond: how ADAMTS13 recognises and cleaves von Willebrand factor. Blood 2011; 118: 3212-3221.
    CrossrefPubMedGoogle Scholar
  • 11 Nolasco LH, Gushiken FC, Turner NA. et al. Protein phosphatase 2B inhibition promotes the secretion of von Willebrand factor from endothelial cells. J Thromb Haemost 2009; 07: 1009-1018.
    CrossrefPubMedGoogle Scholar
  • 12 Dong JF, Moake JL, Nolasco L. et al. ADAMTS13 rapidly cleaves newly secreted ultra-large von Willebrand factor multimers on the endothelial surface under flowing conditions. Blood 2002; 100: 4033-4039.
    CrossrefPubMedGoogle Scholar
  • 13 Nishio K, Anderson PJ, Zheng XL. et al. Binding of platelet GPIb( to von Wille-brand factor domain A stimulates the cleavage of the adjacent domain A2 by ADAMTS13. Proc Natl Acad Sci USA 2004; 101: 10578-10583.
    CrossrefPubMedGoogle Scholar
  • 14 Moake JL, Rudy CK, Troll JH. et al. Unusually large plasma FVIII-von Wille-brand factor multimers in chronic relapsing thrombotic thrombocytopenic purpura. N Engl J Med 1982; 307: 1432-1435.
    CrossrefPubMedGoogle Scholar
  • 15 Furlan M, Robles R, Galbusera M. et al. von Willebrand factor-cleaving protease in acute thrombotic thrombocytopenic purpura. N Engl J Med 1998; 339: 1578-1584.
    CrossrefPubMedGoogle Scholar
  • 16 Tsaai HM, Lian EC. Antibodies to von Willebrand factor cleaving protease in acute thrombotic thrombocytopenic purpura. N Engl J Med 1998; 339: 1585-1594.
    CrossrefPubMedGoogle Scholar
  • 17 Gralnick HR, Williams SB, McKeown LP. et al. In vitro correction of the abnormal multimeric structure of von Willebrand factor in type IIA von Willebrand’s disease. Proc Natl Acad Sci USA 1985; 82: 5968-5972.
    CrossrefPubMedGoogle Scholar
  • 18 Ai J, Smith P, Wang S. et al. The proximal carboxyl-terminal domains of ADAMTS-13 determine substrate specificity and are all required for cleavage of von Willebrand factor. J Biol Chem 2005; 280: 29428-29434.
    CrossrefPubMedGoogle Scholar
  • 19 Gao W, Anderson PJ, Majerus EM. et al. Exosite interactions contribute to tension-induced cleavage of von Willebrand factor by the antithrombotic ADAMTS3 metalloprotease. Proc Natl Acad Sci USA 2006; 103: 19099-19104.
    CrossrefPubMedGoogle Scholar
  • 20 Weiqiang G, Anderson PJ, Sadler JE. Extensive contacts between ADAMTS13 exosites and von Willebrand factor domain A2 contribute to substrate specificity. Blood 2008; 112: 17131719.
    PubMedGoogle Scholar
  • 21 Zanardelli S, Chion ACK, Groot E. et al. A novel binding site for ADAMTS13 constitutively exposed on the surface of globular VWF. Blood 2009; 114: 2819-2828.
    CrossrefPubMedGoogle Scholar
  • 22 Casonato A, De Marco L, Gallinaro L. et al. Altered von Willebrand factor proteolysis and multimer processing associated with the Cys2362Phe mutation in B2 domain. Thromb Haemost 2007; 97: 527-533.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 23 Casonato A, De Marco L, Mazzuccato M. et al. A new congenital platelet abnormality characterized by spontaneous platelet aggregation, enhanced von Wille-brand factor platelet interaction and the presence of all von Willebrand factor multimers in plasma. Blood 1989; 74: 2028-2033.
    PubMedGoogle Scholar
  • 24 Gallinaro L, Cattini MG, Sztukowska M. et al. A shorter von Willebrand factor survival in O blood group subjects explains how ABO determinants influence plasma von Willebrand factor. Blood 2008; 111: 3540-3545.
    CrossrefPubMedGoogle Scholar
  • 25 Newman J, Johnson AJ, Karpatkin MH. et al. Methods for the production of clinically effective intermediate- and high purity factor-VIII concentrates. Br J Haematol 1971; 21: 1-20.
    CrossrefPubMedGoogle Scholar
  • 26 Donadelli R, Orje JN, Capoferri C. et al. Size regulation of von Willebrand factor-mediated platelet thrombi by ADAMTS13 in flowing blood. Blood 2006; 107: 1943-1950.
    CrossrefPubMedGoogle Scholar
  • 27 Morpurgo M, Facchin S, Pignatto M. et al. Characterization of multifunctional nanosystems based on the avidin-nucleic acid interaction as signal enhancers in immuno-detection. Anal Chem 2012; 84: 3433-3439.
    CrossrefPubMedGoogle Scholar
  • 28 Bowen DJ. An influence of ABO blood group on the rate of proteolysis of von Willebrand factor by ADAMTS13. J Thromb Haemost 2003; 01: 33-40.
    CrossrefPubMedGoogle Scholar
  • 29 Eikenboom JC, Castaman G, Vos HL. et al. Characterization of the genetic defects in recessive type 1 and type 3 von Willebrand disease patients of Italian origin. Thromb Haemost 1998; 09: 709-717.
    PubMedGoogle Scholar
  • 30 Tjernberg P, Castaman G, Vos HL. et al. Homozygous C2362F von Willebrand factor induces intracellular retention of mutant von Willebrand factor resulting in autosomal recessive severe von Willebrand disease. Br J Haematol 2006; 133: 409-418.
    CrossrefPubMedGoogle Scholar
  • 31 Castaman G, Eikenboom JC, Lattuada A. et al. Heightened proteolysis of the von Willebrand factor subunit in patients with von Willebrand disease hemizygous or homozygous for the C2362F mutation. Br J Haematol 2000; 108: 188-190.
    CrossrefPubMedGoogle Scholar