Acylation of gelatin-agarose and the enhancement by heparin of fibronectin binding.
The enhancement of the binding of plasma fibronectin to collagen or gelatin by heparin was previously thought to be due primarily to interaction of heparin with fibronectin. We observed, however, that the elution of purified human plasma fibronectin from heparin-treated gelatin-agarose required the same high urea concentrations regardless of whether heparin treatment preceded or followed fibronectin adsorption. Acylation of gelatin-agarose with acetic anhydride or succinic anhydride had little effect upon fibronectin binding, yet the heparin enhancement of fibronectin binding was abolished by either acylation reaction. When heparin binding to gelatin-agarose was investigated with dansyl heparin, gelatin-agarose bound substantial quantities of labeled heparin which could be readily dissociated from the matrix with 2 M NaCl. Acetylated gelatin-agarose did not bind detectable amounts of dansyl heparin. We interpret these results as evidence that the stronger binding of fibronectin to gelatin-agarose in the presence of heparin is due to heparin itself binding to gelatin, thus allowing fibronectin to bind simultaneously to both immobilized ligands through appropriate domains of the glycoprotein.
Smith
R L
RL
Department of Biochemistry and Molecular Biology, Louisiana State University School of Medicine, Shreveport 71130-3932.
Griffin
C A
CA
D000215:Acylation; D002846:Chromatography, Affinity; D005353:Fibronectins / Q000378:metabolism*; D005780:Gelatin / Q000378:metabolism*; D006493:Heparin / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D011485:Protein Binding; D012685:Sepharose; D013329:Structure-Activity Relationship
D005353:Fibronectins; D005780:Gelatin; D006493:Heparin; D012685:Sepharose