The heparin-dependent accumulation of plasma fibronectin on macrophages.
Previous experiments (Hörmann, H. & Jelinić, V. (1980) Hoppe-Seyler's Z. Physiol. Chem. 361, 379-387) had shown that heparin promoted the binding of plasma fibronectin to peritoneal macrophages of guinea pigs. The present data reveal that this effect only takes place at higher fibronectin concentrations indicating cooperative processes, most likely association of fibronectin at the cell surface. An unspecific precipitation of fibronectin by heparin was prevented by calcium in the medium. The accumulation at the cell surface was inhibited by the following fibronectin fragments: N-terminal 30 kDa and 70 kDa containing a potential self-association site and a transamidase-reactive site; central 95 kDa which comprised a negatively charged region possibly involved in self-association as well as the so-called alternative cell-binding site, but was lacking the cell-binding Arg-Gly-Asp sequence; heparin-binding 37-kDa and 60-kDa fragments. All these domains and sites, therefore, were potentially important in the assembly process at the cell surface. A peptide comprising the sequence Arg-Gly-Asp was ineffective pointing against an involvement of this fibronectin cell-binding site in the overall process. Macrophages of older animals were less capable of accumulating fibronectin under the reaction conditions. Their capability was improved after preincubation with activated plasma transglutaminase (coagulation factor XIIIa) suggesting that a cell-attached transamidase might be important for the assembly process.
Max-Planck-Institut für Biochemie, Martinsried bei München.
H
H
Hörmann
V
V
Jelinić
H
H
Richter
D000375:Aging / Q000378:metabolism; D000818:Animals; D005033:Ethylmaleimide / Q000494:pharmacology; D005353:Fibronectins / Q000097:blood*; D006168:Guinea Pigs; D006493:Heparin / Q000494:pharmacology*; D066298:In Vitro Techniques; D007457:Iodine Radioisotopes; D008264:Macrophages / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D010455:Peptides / Q000097:blood; D013997:Time Factors; D000637:Transaminases / Q000378:metabolism
D005353:Fibronectins; D007457:Iodine Radioisotopes; D010455:Peptides; D006493:Heparin; D000637:Transaminases; D005033:Ethylmaleimide