Fibronectin exposes different domains after adsorption to a heparinized and an unheparinized poly(vinyl chloride) surface.
The adsorption of fibronectin to poly(vinyl chloride) catheters with end-point attached (EPA) heparin and tridodecylmethylammonium chloride-heparinized poly(vinyl chloride) was compared to that of unheparinized poly(vinyl chloride) using antibodies directed against four different domains of the protein. After perfusion of human plasma on the EPA-heparinized surface, the exposure of the N-terminal 29-kD fragment increased during the first 5 h of perfusion. Also, the exposure of the 30-kD gelatin-binding and 65-kD cell-binding fragments increased with time, but at a lower level. On the unheparinized catheter, low levels of antibodies bound to the different domains, and the binding showed little variation during the 5 h of plasma perfusion, indicating that the fibronectin molecule does not change configuration to a significant extent on this surface after the initial adsorption. When the EPA-heparinized surface was preabsorbed with human fibrinogen before incubation with fibronectin, significantly less of the 29-kD (fibrin-binding) domain was exposed, and the 30-kD domain was not exposed. Exposure of the 31- and 65-kD domains increased after preadsorption of fibrinogen to the surface. Since fibronectin has heparin-binding domains, it adsorbs differently to a heparinized versus an unheparinized surface. This will influence subsequent binding of other proteins to the surface, as well as potential binding of microbes. The use of antibodies to defined domains of the fibronectin molecule provides a powerful tool in studies of configurational changes of fibronectin after adsorption to different surfaces.
Yu
J L
JL
Department of Medical Microbiology, University of Lund, Sweden.
Johansson
S
S
Ljungh
A
A
D000327:Adsorption; D005353:Fibronectins / Q000737:chemistry*; D006493:Heparin / Q000737:chemistry*; D006801:Humans; D011143:Polyvinyl Chloride / Q000737:chemistry*; D011487:Protein Conformation; D013499:Surface Properties
D005353:Fibronectins; D011143:Polyvinyl Chloride; D006493:Heparin