title: Analysis of the mechanism of recognition in the complement alternative pathway using C3b-bound low molecular weight polysaccharides.
abstract: The human complement (C) system recognizes bacterial, fungal and viral activators of the alternative pathway following covalent attachment of the protein C3b to carbohydrates (CHO) on the surface of the organisms. Recognition first manifests itself as a 3- to 10-fold reduction in the affinity of C3b for factor H, a regulatory protein of C. This report describes the use of a fluorimetric assay which is sensitive to the C3b-H interaction to study the characteristics of recognition. Fluid phase C3b covalently bound to CHO (C3b-CHO) was prepared by activating C3 in the presence of the small homopolymers dextran (alpha 1-6 polyglucose) or inulin (beta 1-2 polyfructose). In particulate form both polysaccharides are activators of C. The conjugates exhibited increased resistance to inactivation in the factor H-dependent assays compared to C3b not bound to CHO and to C3b bound to mono- or disaccharides. The dextran-induced restriction of inactivation was partially reversed by treatment of the conjugate with dextranase. C3b-CHO conjugates failed to bind to factor H-Sepharose and when introduced into serum behaved as though C3b was attached to particulate activators of C, suggesting that the fluorimetric assay accurately reports recognition. The results suggest that the recognition site which induces a reduction in the affinity of C3b for factor H is distinct from the thioester site of C3b and can recognize structural features of polysaccharides including size, sialic acid content, and possibly aspects of three-dimensional oligosaccharide structure.
authors: Department of Biochemistry, University of Texas Health Center, Tyler 75710.
authors: M K
authors: MK
authors: Pangburn
mesh_terms: D001665:Binding Sites; D002846:Chromatography, Affinity; D003167:Complement Activation; D003179:Complement C3b / Q000378:metabolism*; D003180:Complement C3b Inactivator Proteins / Q000378:metabolism; D017242:Complement Factor H; D003170:Complement Pathway, Alternative; D003910:Dextranase / Q000494:pharmacology; D003911:Dextrans / Q000378:metabolism; D006801:Humans; D007444:Inulin / Q000494:pharmacology; D008970:Molecular Weight; D011134:Polysaccharides / Q000378:metabolism*
chemical_list: C493495:CFH protein, human; D003180:Complement C3b Inactivator Proteins; D003911:Dextrans; D011134:Polysaccharides; D003179:Complement C3b; D017242:Complement Factor H; D007444:Inulin; D003910:Dextranase