Differences between the binding sites of the complement regulatory proteins DAF, CR1, and factor H on C3 convertases.
Binding studies using purified decay-accelerating factor (DAF), CR1, and Factor H indicate that the primary interaction of DAF with C3 convertases is with the Bb or C2a subunits, whereas CR1 and Factor H interact primarily with the C3b or C4b subunits. The ability of soluble DAF, CR1, or Factor H to decay C3b,Bb bound to zymosan was inhibited by various concentrations of fluid-phase competitors (C3b, Bb, C3b,Bb, C3b,B, C4b, or C4b,C2a) in 0.1% NP-40 at 22 degrees C. The apparent association constants (appKa) for DAF were 0.045, 0.067, 0.91, 0.71, 0.00045, and 0.53 microM-1, respectively. The appKa for CR1 were 0.50, 0.0040, 1, 1, 1, and 1.1 microM-1, respectively. The appKa for Factor H were 4.3, 0.0005, 2.9, 6.3, 0.27, and 0.29 microM-1, respectively. Thus, C3b binds to DAF with a 10-fold lower affinity than to CR1 and a 100-fold lower affinity than to Factor H. The appKa of C3b,Bb for the three proteins were more similar: DAF (0.91 microM-1), CR1 (1 microM-1), and Factor H (2.9 microM-1). DAF binds to Bb with a 50% higher affinity than to C3b, and to C4b,C2a with a 1000-fold higher affinity than to C4b alone. In contrast, CR1 and Factor H bind almost equally well to the C3 convertases and to their noncatalytic subunits. The affinity of DAF for CVF,Bb was similar to its affinity for Bb alone, suggesting that DAF does not recognize conformational determinants unique to Bb in C3 convertases.
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D001665:Binding Sites; D001667:Binding, Competitive; D001779:Blood Coagulation Factors / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism*; D018958:CD55 Antigens; D003166:Complement Activating Enzymes / Q000378:metabolism*; D003167:Complement Activation; D003175:Complement C2 / Q000378:metabolism; D050678:Complement C2a; D050577:Complement C3-C5 Convertases / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism*; D003181:Complement C4 / Q000378:metabolism; D015935:Complement C4b; D011415:Complement Factor B / Q000378:metabolism; D017242:Complement Factor H; D006801:Humans; D011951:Receptors, Complement / Q000378:metabolism*; D017463:Receptors, Complement 3b
D001779:Blood Coagulation Factors; D001798:Blood Proteins; D018958:CD55 Antigens; C493495:CFH protein, human; D003175:Complement C2; D003180:Complement C3b Inactivator Proteins; D003181:Complement C4; D011951:Receptors, Complement; D017463:Receptors, Complement 3b; D003179:Complement C3b; D015935:Complement C4b; D017242:Complement Factor H; D003166:Complement Activating Enzymes; D050577:Complement C3-C5 Convertases; D050678:Complement C2a; D011415:Complement Factor B