Requirements for the binding of human plasma fibronectin to the C1q subunit of the first component of complement.
We have shown previously that [125I]fibronectin (Fn) binds to solid-phase C1q in a dose-dependent manner. When C1r and C1s were added, the binding of Fn to C1q was abolished; removal of C1r and C1s restored Fn binding to C1q. In this report, we have systematically examined the optimal conditions that favor the Fn-C1q interaction. Our studies show that purified native 125I-Fn binds to C1q in a specific, saturable manner. Maximal 125I-Fn binding to C1q and gelatin occurs at low ionic strength (mu = 0.05) and drops sharply as the ionic strength is increased. At mu = 0.20, the binding to C1q is inhibited by 95%, whereas the binding to gelatin is inhibited by 50%. Optimal binding of Fn to C1q and gelatin occurs between pH 5.5 and 7.5, is decreased by 45% at 4 degrees C, and increases with incubation time; saturation of binding occurs in 60 min at 37 degrees C. Scatchard analysis of binding at mu = 0.05 indicates a single class of high affinity binding sites (Kd = 3.7 X 10(-8) M +/- 0.36 SD). The Kd of the reaction when C1q is bound to either plastic or immune complexes is essentially the same, and, in both cases, increases as the ionic strength of the medium is increased. Finally, significant binding of Fn to C1q can be demonstrated at physiologic ionic strength employing either insoluble immune complexes containing C1q or chemically cross-linked C1q.
D000818:Animals; D000936:Antigen-Antibody Complex / Q000378:metabolism; D003166:Complement Activating Enzymes / Q000378:metabolism*; D015922:Complement C1q; D003432:Cross-Linking Reagents / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009994:Osmolar Concentration; D011487:Protein Conformation; D011817:Rabbits; D011956:Receptors, Cell Surface / Q000032:analysis* / Q000378:metabolism; D017469:Receptors, Fibronectin; D013696:Temperature
D000936:Antigen-Antibody Complex; D003432:Cross-Linking Reagents; D011956:Receptors, Cell Surface; D017469:Receptors, Fibronectin; D015922:Complement C1q; D003166:Complement Activating Enzymes