Structural and functional studies of complement inhibitor C4b-binding protein.
C4b-binding protein (C4BP) is a potent inhibitor of the classical pathway of the complement system. This large plasma glycoprotein consists of seven identical alpha-chains and a unique beta-chain held together by disulphide bridges. Both types of subunits are composed almost exclusively of complement control protein domains (CCPs). Using homology-based computer modelling and mutagenesis of recombinant proteins we have localized binding sites for several ligands of C4BP: complement factor C4b, heparin and vitamin K-dependent anticoagulant protein S (PS). We found that C4b requires CCP1-3 of the alpha-chain for binding. The interaction is ionic in nature and mediated by a cluster of positively charged amino acids present on the interface between CCP1 and CCP2 of the alpha-chain. Loss of C4b-binding resulted in a loss of all inhibitory functions of C4BP within the classical pathway of complement. Binding of heparin required CCPs 1-3 of the alpha-chain, with CCP2 being the most important, as well as the cluster of positively charged amino acids involved in binding of C4b. The interaction between C4BP and PS is of very high affinity and conveyed by a cluster of surface exposed hydrophobic amino acids localized on CCP1 of the beta-chain. Furthermore, C4BP is captured on the surface of several pathogens, which may contribute to their serum resistance and pathogenicity. We have localized interaction of C4BP with Neisseria gonorrhoeae, Bordetella pertussis, Streptococcus pyogenes and Escherichia coli to various regions of the alpha-chain.
Blom
A M
AM
Department of Clinical Chemistry, Lund University, The Wallenberg Laboratory, University Hospital Malmö, Sweden. anna.blom@klkemi.mas.lu.se
D000818:Animals; D001665:Binding Sites; D003169:Complement Inactivator Proteins / Q000737:chemistry* / Q000502:physiology*; D006023:Glycoproteins; D006493:Heparin / Q000378:metabolism; D006801:Humans; D008024:Ligands; D008954:Models, Biological; D011485:Protein Binding; D011487:Protein Conformation; D013329:Structure-Activity Relationship
D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D008024:Ligands; D006493:Heparin