The C3/C5 convertase of the alternative pathway of complement: stabilization and restriction of control by lanthanide ions.
The alternative pathway C3 convertase (C3b,Bb) is a Mg-dependent, labile enzyme with a t/2 of 3 min at 37 degrees C and of 14 min at 24 degrees C (at half physiological ionic strength). To stabilize the enzyme, metal ions of the lanthanide series were tested. Formation and decay of the enzyme as well as binding of radiolabeled Factor B, Factor H or properdin to C3b were measured using C3b-bearing sheep erythrocytes (EC3b). Binding of Factor B to EC3b in presence of 40 microM gadolinium (Gd) was two to three times greater than in presence of 1 mM Mg. Binding of Factor H and of properdin to EC3b was partially inhibited by Gd. Although it enhanced Factor B uptake by EC3b, Gd was unable to substitute for Mg in enzyme formation by Factor D and completely inhibited (at 10 microM) Mg-dependent enzyme activation. However, the preformed enzyme was not inhibited by Gd. Instead, exposure of EC3b,Bb to 40-100 microM Gd increased the t/2 at 37 degrees C from 3 min to 12-28 min, and at 24 degrees C from 14 min to 32 min. The slow decay of the enzyme correlated with slow release of Bb. Similar enzyme stabilization was observed using terbium, ytterbium, dysprosium and lanthanum. The Gd stabilized enzyme was also less susceptible to control by Factor H and properdin than the unstabilized enzyme. Furthermore, Gd protected surface bound-C3b from being cleaved by Factor I. The Gd effects were instantaneously reversed upon addition of 10 mM EDTA. Thus, Gd is able to stabilize preformed C3b,Bb and to render the enzyme refractory to control by Factors H and I.
D002413:Cations, Divalent; D003166:Complement Activating Enzymes / Q000378:metabolism*; D003167:Complement Activation; D050577:Complement C3-C5 Convertases / Q000378:metabolism*; D003179:Complement C3b / Q000378:metabolism; D003182:Complement C5 / Q000378:metabolism; D003170:Complement Pathway, Alternative; D004789:Enzyme Activation / Q000187:drug effects; D005682:Gadolinium / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D008674:Metals, Rare Earth / Q000494:pharmacology*
D002413:Cations, Divalent; D003182:Complement C5; D008674:Metals, Rare Earth; D003179:Complement C3b; D005682:Gadolinium; D003166:Complement Activating Enzymes; D050577:Complement C3-C5 Convertases; D008274:Magnesium