Immobilization of soluble fibrin on factor XIIIa-coated polystyrene beads mediated by N-terminal fibronectin fragments. II. Demonstration of covalent adducts of fibrin peptide chains and fibronectin fragments.
Previous experiments had shown that the free N-terminal fibronectin 30-kDa-domain mediates binding of soluble 125I-fibrin to transamidase-coated polystyrene beads (Hörmann et al., Biol. Chem. Hoppe-Seyler 368, 669-674, 1987). Now, the formation of covalent adducts of the N-terminal fragment with fibrin peptide chains is demonstrated. Binding of soluble 125I-fibrin was performed in presence of N-terminal fibronectin 30-kDa or 70-kDa fragments. The material adsorbed was removed from the beads under reducing conditions and analysed by dodecylsulfate gel electrophoresis followed by autoradiography. The 30-kDa fragment gave rise to bands of 80 kDa and 180-200 kDa which were lacking in the products of the 70-kDa compound. Instead, they showed bands at 120 kDa and ca. 280 kDa. Evidently, those bands represented covalent adducts of fibrin peptide chains or their dimers with the 30-kDa or the 70-kDa fragment, respectively. In addition, dimeric gamma-chains and alpha-chain polymers of fibrin were present indicating partial polymerization of bead-attached fibrin.
Hörmann
H
H
Max-Planck-Institut für Biochemie, Martinsried bei München.
Richter
H
H
Jelinić
V
V
D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000378:metabolism*; D005337:Fibrin / Q000302:isolation & purification / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding; D011503:Transglutaminases / Q000378:metabolism*; D014357:Trypsin
D004800:Enzymes, Immobilized; D005353:Fibronectins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D005337:Fibrin; D011503:Transglutaminases; D014357:Trypsin