title: An Arg760Cys mutation in the consensus sequence of the von Willebrand factor propeptide cleavage site is responsible for a new von Willebrand disease variant.
abstract: We describe a von Willebrand disease (VWD) variant characterized by the persistence of von Willebrand factor (VWF) propeptide as a result of a C>T transition at nucleotide 2527 in exon 17 of the VWF gene. This mutation, which was present in the proband and his father, predicts the substitution of Cys for Arg at position 760 of pre-pro-VWF, 4 residues before the propeptide cleavage site belonging to a consensus sequence for substrate recognition by the processing enzyme paired dibasic amino acid-cleaving enzyme (PACE)/furin. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) documented the presence of both processed and unprocessed VWF in the patient's plasma, with unprocessed VWF relatively less represented. The patient's hemostatic phenotype was characterized by a mild decrease in plasma factor VIII (FVIII) and VWF, a decrease in plasma VWF multimers, and a mild reduction in the FVIII binding capacity of VWF. The FVIII binding defect was more pronounced in the proband than in the father because he also inherited the type 2N Arg91Gln mutation from his mother. The persistence of VWF propeptide did not impair VWF synthesis because platelet VWF content was normal, nor did it compromise VWF storage in endothelial cells, because of the normal post-1-deamino-8-D-arginine vasopressin (DDAVP) increase in plasma VWF. Coexpression of wild-type and Arg760Cys VWF into a Furin-producing BHK cell line resulted in decreased VWF secretion and a defect in the FVIII binding capacity of VWF, together with the persistence of VWF propeptide. These findings confirm that a normal consensus sequence for VWF propeptide cleavage and efficient cleavage are required in vivo for normal FVIII binding capacity of VWF.
authors.affiliation: Department of Medical and Surgical Sciences, Second Chair of Internal Medicine, University of Padua, Medical School, Italy. sandra.casonato@unipd.it
authors.forename: Alessandra
authors.initials: A
authors.lastname: Casonato
authors.forename: Francesca
authors.initials: F
authors.lastname: Sartorello
authors.forename: Maria Grazia
authors.initials: MG
authors.lastname: Cattini
authors.forename: Elena
authors.initials: E
authors.lastname: Pontara
authors.forename: Carmen
authors.initials: C
authors.lastname: Soldera
authors.forename: Antonella
authors.initials: A
authors.lastname: Bertomoro
authors.forename: Antonio
authors.initials: A
authors.lastname: Girolami
mesh_terms: D000328:Adult; D001665:Binding Sites / Q000235:genetics; D016384:Consensus Sequence / Q000235:genetics*; D005169:Factor VIII / Q000378:metabolism; D005192:Family Health; D045683:Furin; D014644:Genetic Variation; D006801:Humans; D008297:Male; D008565:Membrane Proteins; D010641:Phenotype; D017354:Point Mutation; D011498:Protein Precursors / Q000235:genetics*; D013381:Subtilisins; D014842:von Willebrand Diseases / Q000145:classification / Q000235:genetics*; D014841:von Willebrand Factor / Q000235:genetics* / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; D011498:Protein Precursors; D014841:von Willebrand Factor; C067003:von Willebrand factor propolypeptide; D005169:Factor VIII; D013381:Subtilisins; C468731:FURIN protein, human; D045683:Furin