title: Effect of complement-protein-C3b density on the binding of complement factor H to surface-bound C3b.
abstract: Various amounts of the activation fragment C3b of the complement (C) protein C3 were coupled to Sepharose 4B by catalysis with the C3 convertase of the alternative pathway of C. The binding of radioactively labelled C proteins B and H (= factor H) to the C3b-carrying particles was assayed. It was found that the relative binding of H, but not of B, fell rapidly with decreasing densities of solid-phase C3b, suggesting a sigmoidal relationship between C3b density and binding of H. To study the phenomenon in more detail, preformed C3b was coupled to activated thiopropyl-Sepharose 6B at various densities. By using this model system, it was shown that the binding of H/unit amount of C3b was positively correlated to C3b density up to a C3b concentration of about 0.5 mg/ml of gel, whereas binding of B was independent of C3b density. The results show that accumulation of high densities of C3b on a surface creates high-affinity binding sites for H. Because H has recently been shown to form dimers in solution, the interaction of dimeric H with neighbouring C3b molecules is a likely explanation for the phenomenon. The C3b density effect may be a regulatory mechanism keeping the activation of the alternative pathway of C on activating surface within reasonable limits.
authors.affiliation: Finnish Red Cross Blood Transfusion Service, Helsinki, Finland.
authors.forename: V
authors.initials: V
authors.lastname: Koistinen
mesh_terms: D050577:Complement C3-C5 Convertases / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism*; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism*; D017242:Complement Factor H; D004789:Enzyme Activation; D007700:Kinetics; D046911:Macromolecular Substances; D011485:Protein Binding; D012685:Sepharose
chemical_list: D003180:Complement C3b Inactivator Proteins; D046911:Macromolecular Substances; D003179:Complement C3b; D017242:Complement Factor H; D012685:Sepharose; D050577:Complement C3-C5 Convertases