The binding of human complement proteins C5, factor B, beta 1H and properdin to complement fragment C3b on zymosan.
The covalent binding of complement fragment C3b to zymosan by the action of the alternative-pathway C3 convertase and the reversible binding of several complement proteins (component C5, factor B, beta 1H and properdin) to C3b on zymosan have been investigated. When C3b is deposited on zymosan after activation by a surface-bound C3 convertase, the C3b molecules are deposited in foci around the C3 convertase site, with an average of 30 C3b molecules per site. The association constants of C5, factor B, beta 1H, and properdin for C3b bound to zymosan have been determined. The association constants ranged from 6.5 x 10(-5) M-1 for factor B to 2.9 x 10(7) M-1 for properdin. An approximate stoichiometry of 1 : 1 for C5, factor B, and properdin binding to C3b has been observed. Curvilinear Scatchard plots were observed for beta 1H binding to C3b, with the maximal extrapolated ratio of beta 1H to C3b of 0.32. Physiological amounts of properdin increase by 7-fold the affinity constant for factor B binding to C3b with no alteration in the stoichiometry. Similarly, physiological amounts of factor B increase the affinity constant of properdin to C3b about 4-fold with only a small measured difference in stoichiometry. Competition binding studies and protein modification suggest that C5, factor B, beta 1H, and properdin each bind to a distinct region on C3b.
D001667:Binding, Competitive; D002729:Chloromercuribenzoates / Q000494:pharmacology; D003179:Complement C3b / Q000378:metabolism*; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism*; D003182:Complement C5 / Q000378:metabolism*; D011415:Complement Factor B / Q000378:metabolism*; D017242:Complement Factor H; D004229:Dithiothreitol / Q000494:pharmacology; D004792:Enzyme Precursors / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D011414:Properdin / Q000378:metabolism*; D011485:Protein Binding / Q000187:drug effects; D015054:Zymosan / Q000378:metabolism*
C493495:CFH protein, human; D002729:Chloromercuribenzoates; D003180:Complement C3b Inactivator Proteins; D003182:Complement C5; D004792:Enzyme Precursors; D011414:Properdin; D003179:Complement C3b; D017242:Complement Factor H; D015054:Zymosan; D011415:Complement Factor B; D004229:Dithiothreitol