title: Type 2M:Milwaukee-1 von Willebrand disease: an in-frame deletion in the Cys509-Cys695 loop of the von Willebrand factor A1 domain causes deficient binding of von Willebrand factor to platelets.
abstract: This report examines the genetic basis of a variant form of moderately severe von Willebrand disease (vWD) characterized by low plasma von Willebrand factor antigen (vWF:Ag) levels and normal multimerization, typical of type 1 vWD, but disproportionately-low agonist-mediated platelet-binding activity. We identified an in-frame deletion in vWF exon 28 in three generations of affected family members, who are heterozygous for this mutation. The deletion of nucleotides 4,173-4,205 results in the loss of amino acids Arg629-Gln639 in the Cys509-Cys695 loop of the A1 domain in mature vWF. The secreted mutant vWF showed a normal multimeric profile but did not bind to platelets in the presence of optimal concentrations of either ristocetin or botrocetin. The mutant vWF also failed to interact with heparin, and with vWF monoclonal antibody AvW3, which blocks the binding of vWF to GPlb. In addition, mutant vWF showed reduced secretion from transfected cells concomitant with increased intracellular levels. These results confirm that the deletion is the genetic defect responsible for the reduced interaction of vWF with platelets. We have designated this new variant type 2M:Milwaukee-1 vWD. Our analysis suggests that the potential frequency of this phenotype in individuals diagnosed with type 1 vWD is about 0.5%.
authors.affiliation: Blood Center of Southeastern Wisconsin, Milwaukee 53201-2178, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Mancuso
authors.forename: P A
authors.initials: PA
authors.lastname: Kroner
authors.forename: P A
authors.initials: PA
authors.lastname: Christopherson
authors.forename: E A
authors.initials: EA
authors.lastname: Vokac
authors.forename: J C
authors.initials: JC
authors.lastname: Gill
authors.forename: R R
authors.initials: RR
authors.lastname: Montgomery
mesh_terms: D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D001792:Blood Platelets / Q000378:metabolism; D003435:Crotalid Venoms / Q000494:pharmacology; D004252:DNA Mutational Analysis; D005260:Female; D006493:Heparin / Q000378:metabolism; D006579:Heterozygote; D006801:Humans; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D010641:Phenotype; D010980:Platelet Membrane Glycoproteins / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012310:Ristocetin / Q000494:pharmacology; D016415:Sequence Alignment; D017384:Sequence Deletion; D017385:Sequence Homology; D014842:von Willebrand Diseases / Q000145:classification / Q000235:genetics*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D003435:Crotalid Venoms; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D011993:Recombinant Fusion Proteins; D014841:von Willebrand Factor; C028254:von Willebrand factor receptor; D012310:Ristocetin; C030961:botrocetin; D006493:Heparin