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Thromb Haemost 2007; 97(04): 527-533
DOI: 10.1160/TH06-11-0647
Blood Coagulation, Fibrinolysis and Cellular Haemostasis
Schattauer GmbH

Altered von Willebrand factor subunit proteolysis and multimer processing associated with the Cys2362Phe mutation in the B2 domain

Alessandra Casonato
1   Department of Medical and Surgical Sciences, Second Chair of Internal Medicine, University of Padua Medical School, Padua, Italy
,
Luigi De Marco
2   Immunotransfusional Service and Clinical Analysis, I.R.C.C.S., C.R.O., Aviano, Pordenone, Italy
,
Lisa Gallinaro
1   Department of Medical and Surgical Sciences, Second Chair of Internal Medicine, University of Padua Medical School, Padua, Italy
,
Maryta Sztukowska
1   Department of Medical and Surgical Sciences, Second Chair of Internal Medicine, University of Padua Medical School, Padua, Italy
,
Mario Mazzuccato
2   Immunotransfusional Service and Clinical Analysis, I.R.C.C.S., C.R.O., Aviano, Pordenone, Italy
,
Monica Battiston
2   Immunotransfusional Service and Clinical Analysis, I.R.C.C.S., C.R.O., Aviano, Pordenone, Italy
,
Antonio Pagnan
1   Department of Medical and Surgical Sciences, Second Chair of Internal Medicine, University of Padua Medical School, Padua, Italy
,
Zaverio M. Ruggeri
3   Department of Molecular and Experimental Medicine, The Scripps Research Institute, La Jolla, California, USA
› Author AffiliationsFinancial support: This work was supported in part by funds from the Telethon Foundation (Rome, Italy), by grants HL31950, HL42846, HL78784 (to ZMR) from the National Institutes of Health, and by a grant from the Agenzia Spaziale Italiana (ASI).
Further Information

Publication History

Received 17 November 2006

Accepted after resubmission 31 January 2007

Publication Date:
24 November 2017 (online)

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Summary

The normal von Willebrand factor (vWF) multimer pattern results from the ADAMTS-13 cleavage of the Tyr1605-Met1606 bond in the A2 domain of vWF. We identified a patient with severe von Willebrand disease (vWD) homozygously carrying a Cys to Phe mutation in position 2362 of vWF with markedly altered vWF multimers and an abnormal proteolytic pattern. The proband’s phenotype was characterized by a marked drop in plasma vWF antigen and ristocetin cofactor activity, and a less pronounced decrease in FVIII. The vWF multimers lacked any triplet structure, replaced by single bands with an atypical mobility, surrounded by a smear, and abnormally large vWF multimers. Analysis of the plasma vWF subunit's composition revealed the 225 kDa mature form and a single 205 kDa fragment, but not the 176 kDa and 140 kDa fragments resulting from cleavage by ADAMTS-13.The 205 kDa fragment was distinctly visible, along with the normal vWF cleavage products, in the patient's parents who were heterozygous for the Cys2362Phe mutation. Their vWF levels were mildly decreased and vWF multimers were organized in triplets, but also demonstrated abnormally large forms and smearing. Our findings indicate that a proper conformation of the B2 domain, which depends on critical Cys residues, may be required for the normal proteolytic processing of vWF multimers.

Keywords

von Willebrand factor - von Willebrand disease - vWF multimers - vWF proteolysis - vWF processing
 

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