title: Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS-13.
abstract: BACKGROUND
  von Willebrand disease (VWD) type 2 is associated with mutations in von Willebrand factor (VWF) that affect its secretion, multimeric pattern, affinity for platelet receptors and clearance of the protein. While increased proteolysis by a disintegrin-like and metalloprotease with thrombospondin type 1 motifs-13 (ADAMTS-13) has been clearly established for VWF type 2A, only little is known about VWF types 2B and 2M in this regard.
  
  
  OBJECTIVE
  Sensitivity of wild-type (WT) and mutated recombinant (r) VWF to proteolysis by ADAMTS-13 was investigated to better understand the role of this process in the pathophysiology of VWD.
  
  
  METHODS
  We used human rADAMTS-13-WT to digest 11 full-length recombinant forms of VWF carrying molecular abnormalities identified in patients with VWD type 2A (E1638K and P1648S), type 2B (InsM1303, R1306W, R1308P and V1314F) and type 2M (G1324A, E1359K, K1362T, R1374H and I1425F).
  
  
  RESULTS
  Using low ionic strength conditions, all mutations induced increased proteolysis of rVWF by rADAMTS-13 as compared with rVWF-WT. The susceptibility of mutants decreased in the following order: type 2A > type 2B > type 2M > rVWF-WT. At physiological salt concentration (150 mm NaCl) the sensitivity of all rVWF to rADAMTS-13 was significantly decreased. However, type 2A and type 2B mutants still exhibited a significantly higher susceptibility to rADAMTS-13 than rVWF-WT, whereas type 2M mutants normalized.
  
  
  CONCLUSIONS
  Type 2M mutants and rVWF-WT exhibit a similar sensitivity to rADAMTS-13-mediated proteolysis, in agreement with the normal multimeric pattern in vivo. In VWD type 2B, the spontaneous binding to platelets and excessive degradation by ADAMTS-13 of VWF high-molecular-weight multimers may account for their clearance from plasma.
authors.affiliation: INSERM U770 and Université Paris-Sud, Faculté de Médecine IFR93, Le Kremlin-Bicêtre, France.
authors.forename: J
authors.initials: J
authors.lastname: Rayes
authors.forename: A
authors.initials: A
authors.lastname: Hommais
authors.forename: P
authors.initials: P
authors.lastname: Legendre
authors.forename: H
authors.initials: H
authors.lastname: Tout
authors.forename: A
authors.initials: A
authors.lastname: Veyradier
authors.forename: B
authors.initials: B
authors.lastname: Obert
authors.forename: A S
authors.initials: AS
authors.lastname: Ribba
authors.forename: J P
authors.initials: JP
authors.lastname: Girma
mesh_terms: D051722:ADAM Proteins / Q000235:genetics / Q000378:metabolism*; D000071120:ADAMTS13 Protein; D019281:Dimerization; D020022:Genetic Predisposition to Disease; D006801:Humans; D020125:Mutation, Missense; D009994:Osmolar Concentration; D014842:von Willebrand Diseases / Q000145:classification / Q000209:etiology / Q000235:genetics*; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D014841:von Willebrand Factor; D051722:ADAM Proteins; D000071120:ADAMTS13 Protein; C099604:ADAMTS13 protein, human