title: Characterization of the fibrinogen binding domain of bacteriophage lysin from Streptococcus mitis.
abstract: The binding of bacteria to human platelets is a likely central mechanism in the pathogenesis of infective endocarditis. Platelet binding by Streptococcus mitis SF100 is mediated in part by a lysin encoded by the lysogenic bacteriophage SM1. In addition to its role in the phage life cycle, lysin mediates the binding of S. mitis to human platelets via its interaction with fibrinogen on the platelet surface. To better define the region of lysin mediating fibrinogen binding, we tested a series of purified lysin truncation variants for their abilities to bind this protein. These studies revealed that the fibrinogen binding domain of lysin is contained within the region spanned by amino acid residues 102 to 198 (lysin(102-198)). This region has no sequence homology to other known fibrinogen binding proteins. Lysin(102-198) bound fibrinogen comparably to full-length lysin and with the same selectivity for the fibrinogen Aα and Bβ chains. Lysin(102-198) also inhibited the binding in vitro of S. mitis to human fibrinogen and platelets. When assessed by platelet aggregometry, the disruption of the lysin gene in SF100 resulted in a significantly longer time to the onset of aggregation of human platelets than that of the parent strain. The preincubation of platelets with purified lysin(102-198) also delayed the onset of aggregation by SF100. These results indicate that the binding of lysin to fibrinogen is mediated by a specific domain of the phage protein and that this interaction is important for both platelet binding and aggregation by S. mitis.
authors.affiliation: Division of Infectious Diseases, VA Medical Center, 4150 Clement St., San Francisco, CA 94121, USA.
authors.forename: Ho Seong
authors.initials: HS
authors.lastname: Seo
authors.forename: Paul M
authors.initials: PM
authors.lastname: Sullam
mesh_terms: D000595:Amino Acid Sequence; D001422:Bacterial Adhesion; D001792:Blood Platelets / Q000378:metabolism; D026481:Blotting, Far-Western; D004797:Enzyme-Linked Immunosorbent Assay; D005340:Fibrinogen / Q000378:metabolism*; D006801:Humans; D010974:Platelet Aggregation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D017108:Streptococcus Phages / Q000235:genetics / Q000378:metabolism*; D034361:Streptococcus mitis / Q000821:virology*; D014764:Viral Proteins / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D014764:Viral Proteins; D005340:Fibrinogen