Binding of Glycoprotein Srr1 of Streptococcus agalactiae to Fibrinogen Promotes Attachment to Brain Endothelium and the Development of Meningitis

Abstract

The serine-rich repeat glycoprotein Srr1 of Streptococcus agalactiae (GBS) is thought to be an important adhesin for the pathogenesis of meningitis. Although expression of Srr1 is associated with increased binding to human brain microvascular endothelial cells (hBMEC), the molecular basis for this interaction is not well defined. We now demonstrate that Srr1 contributes to GBS attachment to hBMEC via the direct interaction of its binding region (BR) with human fibrinogen. When assessed by Far Western blotting, Srr1 was the only protein in GBS extracts that bound fibrinogen. Studies using recombinant Srr1-BR and purified fibrinogen in vitro confirmed a direct protein-protein interaction. Srr1-BR binding was localized to amino acids 283–410 of the fibrinogen Aα chain. Structural predictions indicated that the conformation of Srr1-BR is likely to resemble that of SdrG and other related staphylococcal proteins that bind to fibrinogen through a “dock, lock, and latch” mechanism (DLL). Deletion of the predicted latch domain of Srr1-BR abolished the interaction of the BR with fibrinogen. In addition, a mutant GBS strain lacking the latch domain exhibited reduced binding to hBMEC, and was significantly attenuated in an in vivo model of meningitis. These results indicate that Srr1 can bind fibrinogen directly likely through a DLL mechanism, which has not been described for other streptococcal adhesins. This interaction was important for the pathogenesis of GBS central nervous system invasion and subsequent disease progression.

Author Summary

Streptococcus agalactiae (Group B streptococcus, GBS) is a leading cause of meningitis in newborns and infants. This life-threatening infection of the brain and surrounding tissues continues to result in a high incidence of morbidity and mortality, despite antibiotic therapy. A key factor in disease production is the ability of this organism to invade the central nervous system, via the bloodstream. We now report that a GBS surface protein called Srr1 binds fibrinogen, a major protein in human blood. This interaction enhances the attachment of GBS to brain vascular endothelial cells, and contributes to the development of meningitis. A mutation in Srr1 that specifically disrupted binding to fibrinogen significantly reduced GBS attachment to brain endothelium, and markedly reduced virulence in an in vivo model of GBS disease. These studies have identified a new mechanism by which Srr1 contributes to GBS invasion of the central nervous system and may provide a basis for novel therapies targeting Srr1 binding.