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HEMOSTASIS, THROMBOSIS, AND VASCULAR BIOLOGY
From the Molecular Haemostasis Laboratory, Department
of Haematology, The Birmingham Children's Hospital NHS Trust; the
Katharine Dormandy Haemophilia Centre and Haemostasis Unit, Department
of Haematology, Royal Free and University College Medical School,
London; the Division of Haematology, University of Leicester; and the
Department of Haematology, The Royal Liverpool University Hospitals,
Liverpool, United Kingdom.
The 3' end of the VWF gene was screened in the affected
members of 3 different families with type 2A (phenotype IID) von
Willebrand disease (vWD). Exons 49 to 52 of the VWF gene
were amplified and screened for mutations by chemical cleavage mismatch
detection. Mismatched bands were detected in exon 52 of 2 patients and
in exon 51 of a third patient. Using direct DNA sequencing, a
heterozygous G8562A transition leading to a Cys2008Tyr substitution was
found in all the patients in family 1, and a T8561A transversion
leading to a Cys2008Ser substitution was found in both patients from
family 2. In a patient from a third family, an 8-base deletion from
nucleotide 8437 to 8444 was identified in exon 51. The 2 mutations in
exon 52 were reproduced by in vitro site-directed mutagenesis of
full-length von Willebrand factor (vWF) cDNA and transiently expressed
in COS-7 cells. The corresponding recombinant VWFs for these 2 mutations exhibited the typical aberrant vWF:Ag multimer pattern seen
in the plasma of the patients. These 3 mutations demonstrate the importance of other carboxy-terminal cysteines in addition to the
reported Cys2010 residue, in the normal dimerization of vWF, and their
essential role in the assembly of normal multimeric vWF. This article has been cited by other articles:
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| Copyright © 2001 by American Society of Hematology Online ISSN: 1528-0020 | |||||||||
