title: Protein A is the von Willebrand factor binding protein on Staphylococcus aureus.
abstract: Endovascular infection is a highly critical complication of invasive Staphylococcus aureus disease. For colonization, staphylococci must first adhere to adhesive endovascular foci. Von Willebrand factor (vWF) is a large, multimeric glycoprotein mediating platelet adhesion at sites of endothelial damage. Earlier it was demonstrated that vWF binds to and promotes the surface adhesion of S. aureus, prompting this effort to identify the vWF adhesin. In Western ligand assays of S. aureus lysates, staphylococcal protein A (SPA) was recognized by purified vWF. Surface plasmon resonance demonstrated the binding of soluble vWF to immobilized recombinant protein A with a K(d) of 1.49 x 10(-8) mol/L. Using flow cytometry, the binding of fluorescein isothiocyanate-labeled vWF to S. aureus was found to be saturable and inhibitable by unlabeled vWF, antiprotein-A antibodies, or IgG. Isogenic Deltaspa::Tc(r) mutants were constructed by the insertion of a tetracycline resistance cassette into spa using allelic replacement, and it exhibited decreased binding of soluble vWF and decreased adhesion to vWF-adsorbed surfaces. The interaction was restored on complementation of the mutants with spa-containing plasmid pSPA7235. In conclusion, protein A confers interaction of S. aureus with soluble and immobilized vWF in a newly discovered function characterizing protein A as a novel member of the staphylococcal surface protein adhesin superfamily and suggesting its potential role in the pathogenesis of endovascular staphylococcal disease.
authors.affiliation: Departments of Anaesthesiology and Surgical Intensive Care Medicine, Division of Experimental and Clinical Hemostasis, Institute of Medical Microbiology, University of Münster, Münster, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Hartleib
authors.forename: N
authors.initials: N
authors.lastname: Köhler
authors.forename: R B
authors.initials: RB
authors.lastname: Dickinson
authors.forename: G S
authors.initials: GS
authors.lastname: Chhatwal
authors.forename: J J
authors.initials: JJ
authors.lastname: Sixma
authors.forename: O M
authors.initials: OM
authors.lastname: Hartford
authors.forename: T J
authors.initials: TJ
authors.lastname: Foster
authors.forename: G
authors.initials: G
authors.lastname: Peters
authors.forename: B E
authors.initials: BE
authors.lastname: Kehrel
authors.forename: M
authors.initials: M
authors.lastname: Herrmann
mesh_terms: D001422:Bacterial Adhesion; D006801:Humans; D009154:Mutation; D011485:Protein Binding; D013203:Staphylococcal Infections / Q000378:metabolism / Q000382:microbiology; D013205:Staphylococcal Protein A / Q000235:genetics / Q000378:metabolism*; D013211:Staphylococcus aureus / Q000378:metabolism*; D014841:von Willebrand Factor / Q000378:metabolism*
chemical_list: D013205:Staphylococcal Protein A; D014841:von Willebrand Factor