Identification of factor-XIIIa-reactive glutaminyl residues in the propolypeptide of bovine von Willebrand factor

J. Takagi, T. Aoyama, Shoko Ueki, H. Ohba, Y. Saito

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

Von Willebrand factor is a large multimeric plasma protein which plays important roles in platelet aggregation, blood coagulation and probably also in the adhesion of endothelial cells. A 100-kDa propeptide, called the propolypeptide of von Willebrand factor (pp-vWF), is generated during biosynthesis. We found that pp-vWF served as a substrate for transglutaminases including human factor XIIIa and guinea pig liver transglutaminase [Usui, T., Takagi, J. and Saito, Y. (1993) J. Biol. Chem. 265, 12311-12316]. As such, it could form cross-linked copolymers with the extracellular matrix protein, laminin, making it all the more likely that pp-vWF plays a role in cell adhesion phenomena [Takagi, J., Sudo, Y., Saito, T. and Saito, Y. (1994) Eur: J. Biochem. 222, 861-867]. In this work, we identified the Gln residues in pp-vWF specifically reacting with blood coagulation factor XIIIa as amine accepters. The fluorescent amine, dansylcadaverine, was employed for labeling the enzyme-reactive sites of the protein. Following partial proteolysis, fragments containing the labeled Gln residues were isolated by passage through an anti-dansyl affinity chromatographic column. Amino acid sequence analyses of the fragments revealed that, out of about 40 Gln residues in pp-vWF, only four could be modified in the factor-XIIIa-catalyzed reaction.

Original languageEnglish
Pages (from-to)773-777
Number of pages5
JournalEuropean Journal of Biochemistry
Volume232
Issue number3
DOIs
Publication statusPublished - 1995
Externally publishedYes

Fingerprint

Factor XIIIa
von Willebrand Factor
Transglutaminases
Amines
Proteolysis
Blood Coagulation Factors
Extracellular Matrix Proteins
Biosynthesis
Cell adhesion
Protein Sequence Analysis
Endothelial cells
Blood Coagulation
Laminin
Platelets
Human engineering
Coagulation
Platelet Aggregation
Cell Adhesion
Liver
Labeling

Keywords

  • Dansylcadaverine
  • Factor XIIIa
  • Propolypeptide
  • Transglutaminase
  • Von Willebrand factor

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of factor-XIIIa-reactive glutaminyl residues in the propolypeptide of bovine von Willebrand factor. / Takagi, J.; Aoyama, T.; Ueki, Shoko; Ohba, H.; Saito, Y.

In: European Journal of Biochemistry, Vol. 232, No. 3, 1995, p. 773-777.

Research output: Contribution to journalArticle

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