Pneumococcal serine-rich repeat protein (PsrP) is a putative adhesin encoded in the Streptococcus pneumoniae pathogenicity island psrP-secY2A2. Challenge of mice with serotype 4, strain TIGR4, and the isogenic mutants T4DeltapsrP and T4DeltapsrP-secY2A2 determined that PsrP was required for bacterial persistence in the lungs but not for colonization in the nasopharynx or replication in the bloodstream during sepsis. In vitro experiments corroborated this anatomical site-specific role; psrP mutants failed to bind to A549 and LA-4 lung cells, yet adhered normally to human nasopharyngeal epithelial cells and to cells from human and rodent capillary endothelial cell lines. We determined that the amino terminus of PsrP mediated adhesion. Microspheres coated with recombinant PsrP(SRR1-BR) (rPsrP(SRR1-BR)) adhered to A549 cells, and moreover, preincubation of cells with rPsrP(SRR1-BR) inhibited TIGR4 adhesion in vitro. Antibodies against rPsrP(SRR1-BR) also neutralized PsrP function; antiserum against rPsrP(SRR1-BR) blocked TIGR4 adhesion in vitro and, following passive immunization, it protected mice against challenge. We conclude that PsrP is an adhesin required for bacterial persistence in the lungs and that rPsrP(SRR1-BR) is a protective antigen.