title: Decay-accelerating factor (DAF), complement receptor 1 (CR1), and factor H dissociate the complement AP C3 convertase (C3bBb) via sites on the type A domain of Bb.
abstract: The AP C3 convertase, C3bBb(Mg(2+)), is subject to irreversible dissociation (decay acceleration) by three proteins: DAF, CR1, and factor H. We have begun to map the factor B (fB) sites critical to these interactions. We generated a panel of fB mutations, focusing on the type A domain because it carries divalent cation and C3b-binding elements. C3bBb complexes were assembled with the mutants and subjected to decay acceleration. Two critical fB sites were identified with a structural model. 1) Several mutations centered at adjacent alpha helices 4 and 5 (Gln-335, Tyr-338, Ser-339, Asp-382) caused substantial resistance to DAF and CR1-mediated decay acceleration but not factor H. 2) Several mutations centered at the alpha 1 helix and adjoining loops (especially D254G) caused resistance to decay acceleration mediated by all three regulators and also increased C3b-binding affinity and C3bBb stability. In the simplest interpretation of these results, DAF and CR1 directly interact with C3bBb at alpha 4/5; factor H likely interacts at some other location, possibly on the C3b subunit. Mutations at the C3b.Bb interface interfere with the normal dissociation of C3b from Bb, whether it is spontaneous or promoted by DAF, CR1, or factor H.
authors.affiliation: Washington University School of Medicine, Department of Medicine, Division of Rheumatology, St. Louis, Missouri 63110, USA. dhourcad@im.wustl.edu
authors.forename: Dennis E
authors.initials: DE
authors.lastname: Hourcade
authors.forename: Lynne
authors.initials: L
authors.lastname: Mitchell
authors.forename: Lisa A
authors.initials: LA
authors.lastname: Kuttner-Kondo
authors.forename: John P
authors.initials: JP
authors.lastname: Atkinson
authors.forename: M Edward
authors.initials: ME
authors.lastname: Medof
mesh_terms: D018958:CD55 Antigens / Q000378:metabolism*; D002460:Cell Line; D050577:Complement C3-C5 Convertases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017242:Complement Factor H / Q000378:metabolism*; D016895:Culture Media, Serum-Free; D006801:Humans; D008958:Models, Molecular; D009097:Multienzyme Complexes; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D011951:Receptors, Complement / Q000378:metabolism*
chemical_list: D018958:CD55 Antigens; C493495:CFH protein, human; D016895:Culture Media, Serum-Free; D009097:Multienzyme Complexes; D011951:Receptors, Complement; D017242:Complement Factor H; D050577:Complement C3-C5 Convertases