Interaction of gelatin with a fluorescein-labeled 42-kDa chymotryptic fragment of fibronectin.
A 42-kDa gelatin binding fragment of human plasma fibronectin was labeled with fluorescein and its fluid-phase interaction with gelatin was investigated. At 25 degrees C in 0.1 M Tris, 0.15 M NaCl, pH 7.3, a dissociation constant, Kd = 0.6 microM, was obtained from the dependence of fluorescence polarization on gelatin concentration. An identical value was obtained for the unlabeled fragment by competition. Binding was unaffected by higher concentrations of NaCl up to 1.0 M, but increased as much as 20-fold at low ionic strength. The dependence of Kd on temperature revealed that dissociation of the complex is accompanied by an increase in entropy. Thus, the interaction is not dominated by either hydrophobic or electrostatic forces; an important role for hydrogen binding is proposed.
D002918:Chymotrypsin / Q000378:metabolism*; D005353:Fibronectins / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005780:Gelatin / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008970:Molecular Weight; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000378:metabolism*; D011485:Protein Binding; D013816:Thermodynamics; D013861:Thiocyanates
D005353:Fibronectins; D005452:Fluoresceins; D005456:Fluorescent Dyes; D010446:Peptide Fragments; D013861:Thiocyanates; D005780:Gelatin; D002918:Chymotrypsin; D016650:Fluorescein-5-isothiocyanate