1a). The threshold for considering a positive interaction was twice the BSA negative control. Consequently, no significant binding of R6 bacteria was detected to collagen type IV, to a mix of different collagens or to elastin. A low binding level (two to three times above the BSA binding level) was observed EVP4593 cost for CRP, fibrinogen, fibronectin, mucin and SAP while a higher level of binding was detected to laminin, lactoferrin, plasminogen and factor H (Fig. 1a). A similar experiment has been performed with the Ruboxistaurin cell line encapsulated TIGR4 strain (Fig 1b). No, or very low binding level,

was observed for the TIGR4 strain to the collagen type IV, fibronectin, mucin and SAP and a slight higher interaction with CRP, fibrinogen, laminin, collagens and elastin. A high binding level of the TIGR4 strain was measured to lactoferrin, plasminogen and factor H (Fig 1b). Both R6 and TIGR4 strains bind strongly to the lactoferrin and factor

H, while the high binding level of R6 to laminin and plasminogen is less important in the selleck kinase inhibitor case of the TIGR4 strain, the latter harbors a higher recognition property to the elastin compared to the R6 strain. Figure 1 Streptococcus pneumoniae interaction with mammalian proteins. FITC labeled bacteria from the R6 and TIGR4 strains were tested for their interaction with several components of the host, extracellular matrix component, circulating proteins or immunity related proteins. BSA is used as a negative control. One representative experiment is

presented in each case. (a) R6 binding pattern. Error bars correspond to the standard deviation of quadruplicates within each sample. (b) Comparison of TIGR4 and R6 and binding Mirabegron pattern. The relative values (residual BSA blank subtracted) are presented for comprehensive comparison of the binding patterns. Interaction of pneumococcal cells with laminin [31], CRP [32], fibronectin [33] and mucin [34] have been described in the literature. All other identified interactions are not described to date, and to investigate these interactions at the molecular level, we designed an approach to systematically test interactions between selected pneumococcal surface proteins and host proteins. Identification, expression and purification of choline-binding proteins (Cbps) We built a list of the Cbps present in the R6 and TIGR4 genomes using the published data [28, 29]. From these sequences, 10 genes encoding Cbps were predicted in the R6 genome, and 15 in the TIGR4 genome (Fig 2). We systematically compared the TIGR4 and R6 protein databases derived from their complete genome sequence in order to get a list of orthologs between the two organisms. This work was facilitated by the high level of conservation of gene organization between both genomes. This analysis led to the identification of two new Cbps in the R6 genome not identified in the initial study [29], namely spr0583 and spr1274 (Fig 2).