title: A novel von Willebrand disease-causing mutation (Arg273Trp) in the von Willebrand factor propeptide that results in defective multimerization and secretion.
abstract: In this report we describe the molecular defect underlying partial and severe quantitative von Willebrand factor (VWF) deficiencies in 3 families previously diagnosed with types 1 and 3 Von Willebrand-disease. Analysis of the VWF gene in affected family members revealed a novel C to T transition at nucleotide 1067 of the VWF complemetary DNA (cDNA), predicting substitution of arginine by tryptophan at amino acid position 273 (R273W) of pre-pro-VWF. Two patients, homozygous for the R273W mutation, had a partial VWF deficiency (VWF:Ag levels of 0.06 IU/mL and 0.09 IU/mL) and lacked high-molecular weight VWF multimers in plasma. A third patient, also homozygous for the R273W mutation, had a severe VWF deficiency (VWF:Ag level of less than 0.01 IU/mL) and undetectable VWF multimers in plasma. Recombinant VWF having the R273W mutation was expressed in COS-7 cells. Pulse-chase experiments showed that secretion of rVWFR273W was severely impaired compared with wild-type rVWF. However, the mutation did not affect the ability of VWF to form dimers in the endoplasmic reticulum (ER). Multimer analysis showed that rVWFR273W failed to form high-molecular-weight multimers present in wild-type rVWF. We concluded that the R273W mutation is responsible for the quantitative VWF deficiencies and aberrant multimer patterns observed in the affected family members. To identify factors that may function in the intracellular retention of rVWFR273W, we investigated the interactions of VWF expressed in COS-7 cells with molecular chaperones of the ER. The R273W mutation did not affect the ability of VWF to bind to BiP, Grp94, ERp72, calnexin, and calreticulin in COS-7 cells. (Blood. 2000;96:560-568)
authors.affiliation: Division of Molecular and Genetic Medicine, Royal Hallamshire Hospital, University of Sheffield, UK. simon.allen@sheffield.ac.uk
authors.forename: S
authors.initials: S
authors.lastname: Allen
authors.forename: A M
authors.initials: AM
authors.lastname: Abuzenadah
authors.forename: J
authors.initials: J
authors.lastname: Hinks
authors.forename: J L
authors.initials: JL
authors.lastname: Blagg
authors.forename: T
authors.initials: T
authors.lastname: Gursel
authors.forename: J
authors.initials: J
authors.lastname: Ingerslev
authors.forename: A C
authors.initials: AC
authors.lastname: Goodeve
authors.forename: I R
authors.initials: IR
authors.lastname: Peake
authors.forename: M E
authors.initials: ME
authors.lastname: Daly
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000378:metabolism; D037281:Calnexin; D037282:Calreticulin; D003241:Consanguinity; D019281:Dimerization; D005260:Female; D006720:Homozygote; D006801:Humans; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D011498:Protein Precursors / Q000235:genetics*; D051381:Rats; D011994:Recombinant Proteins; D012261:Ribonucleoproteins / Q000378:metabolism; D014421:Turkey; D014842:von Willebrand Diseases / Q000235:genetics*; D014841:von Willebrand Factor / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D002135:Calcium-Binding Proteins; D037282:Calreticulin; D011498:Protein Precursors; D011994:Recombinant Proteins; D012261:Ribonucleoproteins; D014841:von Willebrand Factor; D037281:Calnexin