title: On the site of C4 deposition upon complement activation via the mannan-binding lectin pathway or the classical pathway.
abstract: The mannan-binding lectin (MBL) pathway and the classical pathway of complement activation are initiated by the binding of the recognition structure of the initiator complexes, MBL and C1q, respectively, to their ligands, i.e. carbohydrate structures or immune complexes. Proenzymes associated with MBL or C1q are then activated and generate C3 convertase through the activation of C4 and C2. The cleavage product of C4, C4b, attaches covalently to nearby hydroxyl or amino groups. The current picture is that C2 must then attach to C4b before being cleaved by the same associated proteases into the enzymatically active fragment, C2b. This suggests a stringent requirement for the deposition of C4b very close to the initiator complex, or indeed onto the initiator complex. We examined the possibility of C4b being bound to the initiator complex by a solid-phase assay, allowing for the selective elution of the initiator complexes, followed by quantification of the C4b being eluted and the C4b remaining on the solid phase. Also, we estimated the generation of complexes between the released initiator complex and C4b. More than 99% of deposited C4b was bound directly to the solid phase rather than to the initiator complex. Our approach cannot answer the question of the whereabouts of the C2 when it is cleaved.
authors.affiliation: Department of Medical Microbiology and Immunology, University of Aarhus, Aarhus, Denmark. mmk@microbiology.au.dk
authors.forename: M
authors.initials: M
authors.lastname: Møller-Kristensen
authors.forename: S
authors.initials: S
authors.lastname: Thiel
authors.forename: A G
authors.initials: AG
authors.lastname: Hansen
authors.forename: J C
authors.initials: JC
authors.lastname: Jensenius
mesh_terms: D001665:Binding Sites; D003167:Complement Activation; D050577:Complement C3-C5 Convertases / Q000378:metabolism; D003181:Complement C4 / Q000378:metabolism*; D003171:Complement Pathway, Classical; D037582:Complement Pathway, Mannose-Binding Lectin; D006801:Humans; D066298:In Vitro Techniques; D046911:Macromolecular Substances; D018448:Models, Immunological
chemical_list: D003181:Complement C4; D046911:Macromolecular Substances; D050577:Complement C3-C5 Convertases