C3 convertase of human complement: enhanced formation and stability of the enzyme generated with nickel instead of magnesium.
We demonstrate that nickel++ (Ni) can replace Mg in the formation of the C3b,Bb enzyme, and that Ni is more efficient in enzyme formation than Mg. C3b-bearing sheep erythrocytes (EC3b) were used to measure radiolabeled Factor B uptake and C3 convertase activity. Up to nine times more Factor B was specifically bound to EC3b in the presence of Ni than in the presence of Mg under identical conditions. To form one effective hemolytic site (1 Z) per EC3b cell with Ni, three times less Factor B, 12 times less Factor D, and 66 times less metal ion were required than when using Mg. The C3b,Bb formed with Ni (C3b,Bb(Ni)) had a 5 to 10 times longer half-life at different temperatures than the enzyme formed with Mg (C3b,Bb(Mg)). Native properdin stabilized C3b,Bb(Ni) to the same extent as C3b,Bb(Mg) (four to five times) and Factor H accelerated similarly the decay of both enzymes. Ni could also replace Mg in the formation of the C4b,2a enzyme. C4b,2a formation was measured by using C1 and C4b-bearing sheep erythrocytes (EAC1,4b) and native C2. To form 1 Z per EAC1,4b cell with Ni, two times less C2 and 60 times less metal ion were required than when using Mg. The half-life of the C4b,2a formed with Ni was two times longer than that formed with Mg. Decay of both C3b,Bb and C4b,2a enzymes formed either with Mg or with Ni was unaffected by EDTA. These results show that Ni is more efficient than Mg in the generation of both enzymes and suggest that Ni binds to the Mg-binding sites of the enzymes with a higher affinity than Mg.
D003166:Complement Activating Enzymes / Q000378:metabolism*; D003167:Complement Activation; D050577:Complement C3-C5 Convertases / Q000378:metabolism*; D006801:Humans; D046911:Macromolecular Substances; D008274:Magnesium / Q000378:metabolism; D009532:Nickel / Q000378:metabolism*; D011485:Protein Binding; D013329:Structure-Activity Relationship
D046911:Macromolecular Substances; D009532:Nickel; D003166:Complement Activating Enzymes; D050577:Complement C3-C5 Convertases; D008274:Magnesium