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Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor.

Publication ,  Journal Article
Dent, JA; Berkowitz, SD; Ware, J; Kasper, CK; Ruggeri, ZM
Published in: Proc Natl Acad Sci U S A
August 1990

Proteolytic cleavage of the von Willebrand factor subunit may be important for processing and/or function of the molecule and is altered in certain subtypes of von Willebrand disease. It results in the generation of two main fragments with apparent molecular masses of 140 kDa and 176 kDa from the 225-kDa subunit. We have now obtained chemical evidence to locate the protease-sensitive bond between residues Tyr-842 and Met-843, a site that appears to reflect the specificity of calcium-dependent neutral proteases (calpains). Antibodies were raised against four synthetic peptides that represented sequences immediately preceding or following or including the cleavage site. One antibody (against the fragment from Ala-837 through Asp-851) reacted only with the intact subunit, and its epitope included the cleavage site. All others reacted specifically with either the 140-kDa or the 176-kDa fragment, demonstrating their origin from a single cleavage. In samples of purified von Willebrand factor from four of five patients with type IIA von Willebrand disease, the anti-peptide antibodies showed markedly decreased reactivity with either the 140-kDa or the 176-kDa fragment, suggesting the existence of distinct molecular abnormalities clustered around the cleavage site. Thus, in the majority of type IIA patients, a common pathogenetic mechanism may lead to the disappearance of the larger multimers as a consequence of structural changes that may expose a sensitive bond to the action of specific proteases. These studies demonstrate the use of anti-peptide antibodies directed at a relevant structural domain for the immunochemical differentiation of normal and mutant molecules.

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Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

August 1990

Volume

87

Issue

16

Start / End Page

6306 / 6310

Location

United States

Related Subject Headings

  • von Willebrand Factor
  • von Willebrand Diseases
  • Polymerase Chain Reaction
  • Oligonucleotide Probes
  • Mutation
  • Molecular Sequence Data
  • Immunoblotting
  • Humans
  • DNA
  • Base Sequence
 

Citation

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Dent, J. A., Berkowitz, S. D., Ware, J., Kasper, C. K., & Ruggeri, Z. M. (1990). Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor. Proc Natl Acad Sci U S A, 87(16), 6306–6310. https://doi.org/10.1073/pnas.87.16.6306
Dent, J. A., S. D. Berkowitz, J. Ware, C. K. Kasper, and Z. M. Ruggeri. “Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor.Proc Natl Acad Sci U S A 87, no. 16 (August 1990): 6306–10. https://doi.org/10.1073/pnas.87.16.6306.
Dent JA, Berkowitz SD, Ware J, Kasper CK, Ruggeri ZM. Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6306–10.
Dent, J. A., et al. “Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor.Proc Natl Acad Sci U S A, vol. 87, no. 16, Aug. 1990, pp. 6306–10. Pubmed, doi:10.1073/pnas.87.16.6306.
Dent JA, Berkowitz SD, Ware J, Kasper CK, Ruggeri ZM. Identification of a cleavage site directing the immunochemical detection of molecular abnormalities in type IIA von Willebrand factor. Proc Natl Acad Sci U S A. 1990 Aug;87(16):6306–6310.
Journal cover image

Published In

Proc Natl Acad Sci U S A

DOI

ISSN

0027-8424

Publication Date

August 1990

Volume

87

Issue

16

Start / End Page

6306 / 6310

Location

United States

Related Subject Headings

  • von Willebrand Factor
  • von Willebrand Diseases
  • Polymerase Chain Reaction
  • Oligonucleotide Probes
  • Mutation
  • Molecular Sequence Data
  • Immunoblotting
  • Humans
  • DNA
  • Base Sequence