Identification of factor-XIIIa-reactive glutaminyl residues in the propolypeptide of bovine von Willebrand factor.
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. & Saito, Y. (1993) J. Biol. Chem. 268, 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. & 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 acceptors. 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.
Takagi
J
J
Department of Biological Sciences, Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, Japan.
Aoyama
T
T
Ueki
S
S
Ohba
H
H
Saito
Y
Y
Lorand
L
L
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000737:chemistry*; D001665:Binding Sites; D002103:Cadaverine / Q000031:analogs & derivatives / Q000276:immunology / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D005453:Fluorescence; D018698:Glutamic Acid / Q000737:chemistry; D008969:Molecular Sequence Data; D017386:Sequence Homology, Amino Acid; D011503:Transglutaminases / Q000737:chemistry* / Q000378:metabolism; D014841:von Willebrand Factor / Q000737:chemistry / Q000276:immunology*
D000911:Antibodies, Monoclonal; D000941:Antigens; C029417:Von Willebrand antigen; D014841:von Willebrand Factor; D018698:Glutamic Acid; D011503:Transglutaminases; C008542:monodansylcadaverine; D002103:Cadaverine