Leu 697--&gt;Val mutation in mature von Willebrand factor is responsible for type IIB von Willebrand disease

Hilbert, Lysiane; Gaucher, Christine; Romeuf, Christophe de; Mh, Horellou; Vink, Tom; Mazurier, Claudine

doi:10.1182/blood.v83.6.1542.bloodjournal8361542

Cited by 22 publications

(46 citation statements)

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“…The multimeric structure of rVWF was analyzed by 0.1% sodium dodecyl sulfate (SDS) 1% agarose gel electrophoresis [25]. Multimers were revealed using an alkaline phosphatase‐conjugated anti‐VWF polyclonal antibody [26]. Multimeric distribution of all rVWF mutants was similar to that of rVWF‐WT (Fig.…”

Section: Methodsmentioning

confidence: 99%

Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS‐13

Rayes

Hommais

Legendre

et al. 2007

Journal of Thrombosis and Haemostasis

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To cite this article: Rayes J, Hommais A, Legendre P, Tout H, Veyradier A, Obert B, Ribba AS, Girma JP. Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS-13. J Thromb Haemost 2007; 5: 321-8.Summary. 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. Objectives: 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.

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Section: Methodsmentioning

confidence: 99%

Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS‐13

Rayes

Hommais

Legendre

et al. 2007

Journal of Thrombosis and Haemostasis

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“…Plasmid pSV‐VWFH, containing full‐length human VWF cDNA, was a kind gift from L. Hilbert and C. Mazurier (Lille, France) [22]. The vector pSV‐VWF‐P1337L was generated as previously described, directly amplifying the patient's genomic DNA in correspondence with the mutation 4010 C > T [23].…”

Section: Methodsmentioning

confidence: 99%

Biochemical characterization of a recombinant von Willebrand factor (VWF) with combined type 2B and type 1 defects in the VWF gene in two patients with a type 2A phenotype of von Willebrand disease

Baronciani

Federici

Cozzi

et al. 2007

Journal of Thrombosis and Haemostasis

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, an infusion test of desmopressin was followed by mild thrombocytopenia. This led to further laboratory investigations of his affected brother and of family members, who showed different phenotypic patterns compatible with type 1, 2A, 2B and an uncertain classification of VWD. The two brothers were compound heterozygotes (C275R/ P1337L), whereas the others members of the family were heterozygous for C275R (a novel mutation in the D1 domain) or P1337L (a type 2B mutation in the A1 domain). Objective and methods: To evaluate the role of the combined effect of the two mutations in the two brothers, C275R and P1337L recombinant (r) VWFs were transiently expressed in COS-7 cells. Results: Recombinant VWF levels secreted in cell media were similar for wild-type (WT), P1337L and hybrid P1337L/ WT rVWFs, reduced for hybrids C275R/P1337L and C275R/ WT rVWFs, and strongly reduced for C275R rVWF. All rVWFs had a full set of multimers except C275R rVWF, which had only dimers. P1337L rVWF and C275R/P1337L rVWF showed the highest degree of binding to glycoprotein (GP) Iba and the lowest to collagen, followed by P1337L/WT rVWF (with an intermediate level of binding to both ligands), and by WT rVWF with the lowest level of binding to GPIba and the highest to collagen. Conclusion: These results suggest that the two compound heterozygous patients have a circulating VWF mainly mutated in the A1 domain (P1337L). This peculiar type 2B VWF variant showed a remarkably high affinity for the GPIba platelet receptor, leading to the loss of high and intermediate molecular weight multimers and hence to decreased RIPA, as in type 2A VWD.

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“…Blood samples were collected into 13 mmol/l sodium citrate, and tests were performed on citrated platelet‐poor plasma samples stored frozen at −20°C. FVIII coagulant activity (FVIII:C), VWF antigen (VWF:Ag) level and ristocetin cofactor (VWF:RCo) activity were assayed as previously described (Hilbert et al , 1994). Measurement of VWF:FVIIIB was carried out in a solid phase system, as previously described (Jorieux et al , 2000).…”

Section: Laboratory Methodsmentioning

confidence: 99%

“…Measurement of VWF:FVIIIB was carried out in a solid phase system, as previously described (Jorieux et al , 2000). Multimeric analysis of VWF was performed by 0·1% sodium dodecyl sulphate (SDS)−1·5% agarose gel electrophoresis followed by visualization of the multimers with alkaline phosphatase‐conjugated immunopurified anti‐VWF polyclonal antibodies (Hilbert et al , 1994).…”

Section: Laboratory Methodsmentioning

confidence: 99%

“…COS‐7 cells were transiently transfected using the diethylaminoethyl‐dextran method (Hilbert et al , 1994). Forty hours after transfection, cells were incubated in serum‐free Dulbecco's modified Eagle medium for 72 h. Then cell‐conditioned media were collected in the presence of 10 mmol/l benzamidine and 1 mmol/l phenylmethylsulfonyl fluoride (PMSF) and cleared from cellular fragments by centrifugation for 5 min at 2500 g .…”

Section: Plasmid Construction and Expressionmentioning

confidence: 99%

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Expression of two type 2N von Willebrand disease mutations identified in exon 18 of von Willebrand factor gene

et al. 2004

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SummaryType 2N von Willebrand disease (VWD) is characterized by a markedly decreased affinity of von Willebrand factor (VWF) for factor VIII (FVIII). The FVIII binding site has been localized within the first 272 amino acid residues of mature VWF, encoded by exons 18-23. Two substitutions in exon 18 of VWF gene, inducing candidate mutations Y795C and C804F were identified in the heterozygous state in two French patients who also displayed the frequent R854Q mutation in exon 20. Expression studies in Cos-7 cells showed that these abnormalities, which implicate cysteine residues, induced secretion, multimerization and FVIII binding defects of corresponding recombinant VWF. Results from transfection experiments with R854Q, performed to reproduce the hybrid VWF present in patient plasma, were in agreement with those obtained for patient's plasma VWF. These findings confirm the importance of the VWF D' domain in FVIII binding. In addition, this work shows that exon 18 should preferentially be sequenced in type 2N VWD patients when the frequent R854Q mutation in exon 20 has been excluded or detected in the heterozygous state.

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Leu 697-->Val mutation in mature von Willebrand factor is responsible for type IIB von Willebrand disease

Cited by 22 publications

References 0 publications

Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS‐13

Effect of von Willebrand disease type 2B and type 2M mutations on the susceptibility of von Willebrand factor to ADAMTS‐13

Biochemical characterization of a recombinant von Willebrand factor (VWF) with combined type 2B and type 1 defects in the VWF gene in two patients with a type 2A phenotype of von Willebrand disease

Expression of two type 2N von Willebrand disease mutations identified in exon 18 of von Willebrand factor gene

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