title: The emerging pathogen Moraxella catarrhalis interacts with complement inhibitor C4b binding protein through ubiquitous surface proteins A1 and A2.
abstract: Moraxella catarrhalis ubiquitous surface protein A2 (UspA2) mediates resistance to the bactericidal activity of normal human serum. In this study, an interaction between the complement fluid phase regulator of the classical pathway, C4b binding protein (C4BP), and M. catarrhalis mutants lacking UspA1 and/or UspA2 was analyzed by flow cytometry and a RIA. Two clinical isolates of M. catarrhalis expressed UspA2 at a higher density than UspA1. The UspA1 mutants showed a decreased C4BP binding (37.6% reduction), whereas the UspA2-deficient Moraxella mutants displayed a strongly reduced (94.6%) C4BP binding compared with the wild type. In addition, experiments with recombinantly expressed UspA1(50-770) and UspA2(30-539) showed that C4BP (range, 1-1000 nM) bound to the two proteins in a dose-dependent manner. The equilibrium constants (K(D)) for the UspA1(50-770) and UspA2(30-539) interactions with a single subunit of C4BP were 13 microM and 1.1 microM, respectively. The main isoform of C4BP contains seven identical alpha-chains and one beta-chain linked together with disulfide bridges, and the alpha-chains contain eight complement control protein (CCP) modules. The UspA1 and A2 bound to the alpha-chain of C4BP, and experiments with C4BP lacking CCP2, CCP5, or CCP7 showed that these three CCPs were important for the Usp binding. Importantly, C4BP bound to the surface of M. catarrhalis retained its cofactor activity as determined by analysis of C4b degradation. Taken together, M. catarrhalis interferes with the classical complement activation pathway by binding C4BP to UspA1 and UspA2.
authors.affiliation: Department of Medical Microbiology, Lund Univerity, Malmö University Hospital, Sweden.
authors.forename: Therése
authors.initials: T
authors.lastname: Nordström
authors.forename: Anna M
authors.initials: AM
authors.lastname: Blom
authors.forename: Arne
authors.initials: A
authors.lastname: Forsgren
authors.forename: Kristian
authors.initials: K
authors.lastname: Riesbeck
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics / Q000276:immunology; D000942:Antigens, Bacterial / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D000954:Antigens, Surface / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D001422:Bacterial Adhesion / Q000235:genetics / Q000276:immunology; D001425:Bacterial Outer Membrane Proteins / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D001770:Blood Bactericidal Activity; D003175:Complement C2 / Q000378:metabolism; D015935:Complement C4b / Q000378:metabolism*; D017244:Complement Factor I / Q000378:metabolism; D003169:Complement Inactivator Proteins / Q000378:metabolism*; D004306:Dose-Response Relationship, Immunologic; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D001936:Moraxella catarrhalis / Q000235:genetics / Q000276:immunology* / Q000472:pathogenicity*; D016254:Mutagenesis, Insertional; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics / Q000276:immunology; D021122:Protein Subunits / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014774:Virulence
chemical_list: D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D003175:Complement C2; D003169:Complement Inactivator Proteins; D006023:Glycoproteins; D010446:Peptide Fragments; D021122:Protein Subunits; D011994:Recombinant Proteins; C089675:UspA protein, Moraxella catarrhalis; D015935:Complement C4b; D017244:Complement Factor I