Regulation of the human alternative complement pathway: formation of a ternary complex between factor H, surface-bound C3b and chemical groups on nonactivating surfaces.
Sephadex [alpha(1----6) cross-linked dextran] activates the human alternative pathway of complement. Substitution of hydroxyl groups of Sephadex with carboxymethyl groups (CM) results in a dose-dependent decrease of the activating capacity of the polymer in normal human serum. Sephadex bearing one CM group/glycosyl unit (CM-Seph 0.95) exhibited no activating capacity. CM groups did not interfere with the ability of the polymer to covalently bind C3b in the presence of purified alternative pathway proteins nor with the capacity of bound-C3b to form a C3 convertase in the absence of regulatory proteins. C3b that was bound to CM-Seph 0.95 was more susceptible to inactivation by factors H and I in serum than C3b bound to Sephadex. Binding studies using 125I-labeled H demonstrated that H bound with a similar affinity to the activating particle Sephadex, to Sephadex bearing C3b and to the nonactivating particle CM-Seph 0.95. However, factor H bound with a 5- to 7-fold higher affinity to CM-Seph 0.95 bearing C3b. These results demonstrate a requirement for both CM groups and C3b molecules in order for H to bind with high affinity to C3b on the non-activating surface, and indicate that H formed a ternary complex with surface-bound C3b and CM groups on CM-Seph 0.95. Using a chemically defined model system, the present study provides a molecular basis for the enhanced interaction between surface-bound C3b and factor H on nonactivators of the human alternative pathway.
Carreno
M P
MP
INSERM U 28, Hôpital Broussais, Paris, France.
Labarre
D
D
Maillet
F
F
Jozefowicz
M
M
Kazatchkine
M D
MD
D003167:Complement Activation; D003179:Complement C3b / Q000378:metabolism*; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism*; D011415:Complement Factor B / Q000378:metabolism; D011416:Complement Factor D / Q000378:metabolism; D017242:Complement Factor H; D003170:Complement Pathway, Alternative; D003911:Dextrans; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D013329:Structure-Activity Relationship
C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D003911:Dextrans; D046911:Macromolecular Substances; D003179:Complement C3b; D017242:Complement Factor H; C025614:sephadex; C493488:CFD protein, human; D011416:Complement Factor D; D011415:Complement Factor B