title: Preferential inactivation of the C5 convertase of the alternative complement pathway by factor I and membrane cofactor protein (MCP).
abstract: Human C3b bound to the ghost of sheep erythrocytes (E*) via activation of the alternative complement pathway (E*AC3b) consists of four major constituents on SDS-PAGE of 350, 260, 210 and 180 kDa. 350 kDa C3b is a dimeric form of C3b in which the alpha' chain of one C3b binds covalently to that of the other C3b. This complex is presumed to serve as a core for the alternative pathway C5 convertase. The other C3b populations are monomers complexed with membrane proteins or sugars. Using E*AC3b (C3b labeled) as a substrate, we have investigated functional properties of membrane cofactor protein (MCP), which is an integral membrane protein with C3b-binding and factor I-dependent cofactor activities. In conjunction with factor I, MCP was found to degrade the protein-bound C3b preferentially including the 350 kDa dimer. There was a similar but lesser tendency of this selective cleavage of C3b-dimer by CR1 but not by factor H or C4bp. In contrast to CR1 and factor H, detergent solubilization of EAC3b was required for MCP to fully express its cofactor activity for this selective degradation of C3b. We next separated the C3b dimer from the monomers and assessed their ability to assemble the alternative C5 convertase. The C3b dimer but not the monomers expressed C5 convertase activity following the addition of factors B and D, C5 and Ni2+. Kinetic analysis of the degradation of the C3b dimer by MCP and factor I suggested that only one C3b was efficiently converted to C3bi and this occurred concomitant with a decrease in C5 convertase activity. These results suggest that MCP has the ability to more efficiently interact with protein-bound C3b and that this may relate as well to its preferential ability to irreversibly inactivate the C5 convertase.
authors.affiliation: Department of Immunology, Center for Adult Diseases Osaka, Japan.
authors.forename: T
authors.initials: T
authors.lastname: Seya
authors.forename: M
authors.initials: M
authors.lastname: Okada
authors.forename: M
authors.initials: M
authors.lastname: Matsumoto
authors.forename: K S
authors.initials: KS
authors.lastname: Hong
authors.forename: T
authors.initials: T
authors.lastname: Kinoshita
authors.forename: J P
authors.initials: JP
authors.lastname: Atkinson
mesh_terms: D015703:Antigens, CD; D050577:Complement C3-C5 Convertases / Q000037:antagonists & inhibitors*; D003179:Complement C3b / Q000378:metabolism*; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D015936:Complement C5a / Q000096:biosynthesis; D017242:Complement Factor H; D017244:Complement Factor I; D003170:Complement Pathway, Alternative; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000378:metabolism*
chemical_list: D015703:Antigens, CD; C497857:CD46 protein, human; C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D011092:Polyethylene Glycols; D003179:Complement C3b; D015936:Complement C5a; D017242:Complement Factor H; D017830:Octoxynol; C010615:Nonidet P-40; D050577:Complement C3-C5 Convertases; D012697:Serine Endopeptidases; D017244:Complement Factor I