title: Transforming growth factor-beta 1 binds to immobilized fibronectin.
abstract: We have characterized the interaction of homodimeric porcine transforming growth factor-beta 1 (TGF-beta 1) with affinity-purified human plasma fibronectin. Using a solid-phase binding assay, we have demonstrated that TGF-beta 1 binds to fibronectin immobilized on Immunlon ITM microtiter plates. TGF-beta 1 binding increased with time, reaching a plateau after 4-6 h, and was dependent upon the concentration of both labeled TGF-beta 1 and immobilized fibronectin present. The binding of radiolabeled TGF-beta 1 to fibronectin was saturable and was reduced 75% in the presence of a 100-fold excess of unlabeled TGF-beta 1. TGF-beta 1 bound to fibronectin with an association rate constant (Ka) of 2.96 x 10(3) M-1 s-1 and did not readily dissociate under various conditions. The binding of TGF-beta 1 to fibronectin was insensitive to variations in ionic strength over a range of 0.1-1.0 M NaCl and was relatively insensitive to divalent cation concentration in the range of 0.1-10.0 mM as well. These data suggest that the binding of TGF-beta 1 to fibronectin may not be dependent upon the interaction of charged amino acids within these two molecules. However, the binding of TGF-beta 1 to fibronectin was strongly pH-dependent and binding decreased dramatically below pH 4.0 and above pH 10.0, suggesting that charged amino acids may influence TGF-beta 1/fibronectin interactions. The association of TGF-beta 1 with immobilized fibronectin or other extracellular matrix components and subsequent dissociation under acidic conditions or by an as-yet-unidentified mechanism may play a role in the distribution and/or activity of this potent growth regulator at sites of tissue injury and inflammation in vivo.
authors.affiliation: Department of Laboratory Medicine and Pathology, University of Minnesota, Minneapolis 55455.
authors.forename: D L
authors.initials: DL
authors.lastname: Mooradian
authors.forename: R C
authors.initials: RC
authors.lastname: Lucas
authors.forename: J A
authors.initials: JA
authors.lastname: Weatherbee
authors.forename: L T
authors.initials: LT
authors.lastname: Furcht
mesh_terms: D000818:Animals; D002118:Calcium / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D005353:Fibronectins / Q000378:metabolism*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009994:Osmolar Concentration; D013552:Swine; D013997:Time Factors; D015291:Transforming Growth Factors / Q000378:metabolism*
chemical_list: D005353:Fibronectins; D015291:Transforming Growth Factors; D006493:Heparin; D002118:Calcium