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

Ho Seong Seo,Paul M Sullam,Brandon J Kim,Rong Mu,Kelly S Doran,Michael R Wessels

doi:10.1371/journal.ppat.1002947

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.

Highlights

The serine-rich repeat (SRR) glycoproteins are a large and diverse family of adhesins found in Gram-positive bacteria [1,2]
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
We report that a GBS surface protein called Srr1 binds fibrinogen, a major protein in human blood

Summary

Introduction

The serine-rich repeat (SRR) glycoproteins are a large and diverse family of adhesins found in Gram-positive bacteria [1,2]. The SRR proteins have a highly conserved domain organization, including a long and specialized signal sequence, two extensive serine-rich repeat regions that undergo glycosylation, and a typical LPXTG cell wall anchoring motif [3,4]. Among the best-characterized is GspB of Streptococcus gordonii, which binds human platelets through its interaction with sialyl-T antigen on the platelet receptor GPIba [2,5]. This appears to be an important event in the pathogenesis of infective endocarditis, since disruption of Siglec-mediated binding results in reduced virulence, as measured by an animal model of endocardial infection [3,4]. PsrP binds cytokeratin 10 in vitro, which appears to be important for binding to pulmonary epithelial cells and subsequent pneumonia [12]

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