title: Trimeric autotransporter DsrA is a major mediator of fibrinogen binding in Haemophilus ducreyi.
abstract: Haemophilus ducreyi is the etiologic agent of the sexually transmitted genital ulcer disease chancroid. In both natural and experimental chancroid, H. ducreyi colocalizes with fibrin at the base of the ulcer. Fibrin is obtained by cleavage of the serum glycoprotein fibrinogen (Fg) by thrombin to initiate formation of the blood clot. Fg binding proteins are critical virulence factors in medically important Gram-positive bacteria. H. ducreyi has previously been shown to bind Fg in an agglutination assay, and the H. ducreyi Fg binding protein FgbA was identified in ligand blotting with denatured proteins. To better characterize the interaction of H. ducreyi with Fg, we examined Fg binding to intact, viable H. ducreyi bacteria and identified a novel Fg binding protein. H. ducreyi bound unlabeled Fg in a dose-dependent manner, as measured by two different methods. In ligand blotting with total denatured cellular proteins, digoxigenin (DIG)-Fg bound only two H. ducreyi proteins, the trimeric autotransporter DsrA and the lectin DltA; however, only the isogenic dsrA mutant had significantly less cell-associated Fg than parental strains in Fg binding assays with intact bacteria. Furthermore, expression of DsrA, but not DltA or an empty vector, rendered the non-Fg-binding H. influenzae strain Rd capable of binding Fg. A 13-amino-acid sequence in the C-terminal section of the passenger domain of DsrA appears to be involved in Fg binding by H. ducreyi. Taken together, these data suggest that the trimeric autotransporter DsrA is a major determinant of Fg binding at the surface of H. ducreyi.
authors.affiliation: Departments of Medicine.
authors.forename: William G
authors.initials: WG
authors.lastname: Fusco
authors.forename: Christopher
authors.initials: C
authors.lastname: Elkins
authors.forename: Isabelle
authors.initials: I
authors.lastname: Leduc
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001770:Blood Bactericidal Activity / Q000276:immunology*; D002352:Carrier Proteins / Q000378:metabolism*; D002602:Chancroid / Q000276:immunology* / Q000378:metabolism; D004076:Digoxigenin / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D006191:Haemophilus ducreyi / Q000276:immunology / Q000378:metabolism; D006801:Humans; D011485:Protein Binding / Q000276:immunology
chemical_list: D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D002352:Carrier Proteins; D005340:Fibrinogen; D004076:Digoxigenin
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