Triplet structure of von Willebrand factor reflects proteolytic degradation of high molecular weight multimers.

Furlan, Miha; Robles, Rodolfo; Affolter, Danilo; Meyer, Dominique; Baillod, Peter; Lämmle, B

doi:10.1073/pnas.90.16.7503

Cited by 80 publications

(55 citation statements)

References 29 publications

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“…The appearance of the triplet structure of vWF is interpreted as a proteolytic product of vWF multimers from the metalloprotease ADAMTS13 (Fernandez et al 1982, Furlan et al 1993). The role of ADAMTS13 in the degradation of vWF has been well established ).…”

Section: Resultsmentioning

confidence: 99%

“…The amount of vWF is normal, but the multimer structure of the vWF is abnormal, or subgroups of large or small multimers are absent. The different subtypes 2M and 2N can be differentiated by more subtle alterations of the inner structure of smaller multimers (Furlan et al 1993;Ruggeri 1999). High-resolution agarose gels resolve the inner multimeric structure so that each smaller multimer dissociates into three distinct subbands.…”

Section: Introductionmentioning

confidence: 99%

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Native multimer analysis of plasma and platelet von Willebrand factor compared to denaturing separation: Implication for the interpretation of satellite bands

et al. 2011

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Blue native electrophoresis (BNE) was applied to analyze the von Willebrand factor (vWF) multimers in their native state and to present a methodology to perform blue native electrophoresis on human plasma proteins, which has not been done before. The major difference between this method and the commonly used SDS‐agarose gel electrophoresis is the lack of satellite bands in the high‐resolution native gel. To further analyze this phenomenon, a second dimension was performed under denaturing conditions. Thereby, we obtained a pattern in which each protein sub‐unit from the first dimension dissociates into three distinct sub‐bands. These bands confirm the triplet structure, which consists of an intermediate band and two satellite bands. By introducing the second dimension, our novel method separates the triplet structure into a higher resolution than the commonly used SDS‐agarose gel electrophoresis does. This helps considerably in the classification of ambiguous von Willebrand's disease subtypes. In addition, our method has the additional advantage of being able to resolve the triplet structure of platelet vWF multimers, which has not been identified previously through conventional SDS‐agarose electrophoresis multimer analysis. This potential enables us to compare the triplet structure from platelet and plasmatic vWF, and may help to find out whether structural abnormalities concern the vWF molecule in the platelet itself, or whether they are due to the physiological processing of vWF shed into circulation. Owing to its resolution and sensitivity, this native separation technique offers a promising tool for the analysis and detection of von Willebrand disorder, and for the classification of von Willebrand's disease subtypes.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Native multimer analysis of plasma and platelet von Willebrand factor compared to denaturing separation: Implication for the interpretation of satellite bands

et al. 2011

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“…8 However, UL multimers do not normally circulate because they are cleaved into smaller multimers soon after their secretion. [10][11][12][13] In patients with TTP or HUS, in contrast to healthy subjects, UL multimers are sometimes detected in plasma. 14,15 The relationship between UL VWF multimers and thrombotic microangiopathies remained elusive until the recent demonstration of the presence in human plasma of a metalloprotease (ADAMTS13) [16][17][18] that cleaves VWF physiologically at the peptide bond between amino acid residues 842Thr and 843Met in the A2 domain of the subunit.…”

Section: Introductionmentioning

confidence: 99%

von Willebrand factor cleaving protease (ADAMTS13) is deficient in recurrent and familial thrombotic thrombocytopenic purpura and hemolytic uremic syndrome

Remuzzi

Galbusera

Noris

et al. 2002

Blood

184

153

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for the Italian Registry of Recurrent and Familial HUS/TTP Whether measurement of ADAMTS13 activity may enable physicians to distinguish thrombotic thrombocytopenic purpura (TTP) from hemolytic uremic syndrome (HUS) is still a controversial issue. Our aim was to clarify whether patients with normal or deficient ADAMTS13 activity could be distinguished in terms of disease manifestations and multimeric patterns of plasma von Willebrand factor (VWF). ADAMTS13 activity, VWF antigen, and multimeric pattern were evaluated in patients with recurrent and familial TTP (n ‫؍‬ 20) and HUS (n ‫؍‬ 29). Results of the collagen-binding assay of ADAMTS13 activity were confirmed in selected samples by testing the capacity of plasma to cleave recombinant VWF A1-A2-A3. Most patients with TTP had complete or partial deficiency of ADAMTS13 activity during the acute phase, and in some the defect persisted at remission. However, complete ADAMTS13 deficiency was also found in 5 of 9 patients with HUS during the acute phase and in 5 patients during remission. HUS patients with ADAMTS13 deficiency could not be distinguished clinically from those with normal ADAMTS13. In a subgroup of patients with TTP or HUS, the ADAMTS13 defect was inherited, as documented by half-normal levels of ADAMTS13 in their asymptomatic parents, consistent with the heterozygous carrier state. In patients with TTP and HUS there was indirect evidence of increased VWF fragmentation, and this occurred also in patients with ADAMTS13 deficiency. In conclusion, deficient ADAMTS13 activity does not distinguish TTP from HUS, at least in the recurrent and familial forms, and it is not the only determinant of VWF abnormalities in these conditions. (Blood. 2002;100:778-785)

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“…Proteolysis of pd-vWF may also result in loss of the largest, r lost hemostatically active cWF multimers [7,17]. However, t-oth r-vWF1808 and r-vWF1904 exhibited very high molecular v eight multimers absent in pd-vWF (Fig.…”

Section: Resultsmentioning

confidence: 99%

Structural analysis of recombinant von Willebrand factor produced at industrial scale fermentation of transformed CHO cells co‐expressing recombinant furin

Fischer¹,

Schlokat²,

Mitterer³

et al. 1995

FEBS Letters

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Triplet structure of von Willebrand factor reflects proteolytic degradation of high molecular weight multimers.

Cited by 80 publications

References 29 publications

Native multimer analysis of plasma and platelet von Willebrand factor compared to denaturing separation: Implication for the interpretation of satellite bands

Native multimer analysis of plasma and platelet von Willebrand factor compared to denaturing separation: Implication for the interpretation of satellite bands

von Willebrand factor cleaving protease (ADAMTS13) is deficient in recurrent and familial thrombotic thrombocytopenic purpura and hemolytic uremic syndrome

Structural analysis of recombinant von Willebrand factor produced at industrial scale fermentation of transformed CHO cells co‐expressing recombinant furin

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