title: Streptococcus agalactiae CspA is a serine protease that inactivates chemokines.
abstract: Streptococcus agalactiae (group B Streptococcus [GBS]) remains a leading cause of invasive infections in neonates and has emerged as a pathogen of the immunocompromised and elderly populations. The virulence mechanisms of GBS are relatively understudied and are still poorly understood. Previous evidence indicated that the GBS cspA gene is necessary for full virulence and the cleavage of fibrinogen. The predicted cspA product displays homology to members of the extracellular cell envelope protease family. CXC chemokines, many of which can recruit neutrophils to sites of infection, are important signaling peptides of the immune system. In this study, we purified CspA and demonstrated that it readily cleaved the CXC chemokines GRO-alpha, GRO-beta, GRO-gamma, neutrophil-activating peptide 2 (NAP-2), and granulocyte chemotactic protein 2 (GCP-2) but did not cleave interleukin-8. CspA did not cleave a panel of other test substrates, suggesting that it possesses a certain degree of specificity. CXC chemokines also underwent cleavage by whole GBS cells in a cspA-dependent manner. CspA abolished the abilities of three representative CXC chemokines, GRO-gamma, NAP-2, and GCP-2, to attract and activate neutrophils. Genetic and biochemical evidence indicated that CspA is a serine protease with S575 at its active site. D180 was also implicated as part of the signature serine protease catalytic triad, and both S575 and D180 were required for both N-terminal and C-terminal autocatalytic processing of CspA.
authors.affiliation: Department of Microbiology and Immunology, LSU Health Sciences Center in Shreveport, Shreveport, Louisiana 71130, USA.
authors.forename: Joshua D
authors.initials: JD
authors.lastname: Bryan
authors.forename: Daniel W
authors.initials: DW
authors.lastname: Shelver
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D020134:Catalytic Domain; D002478:Cells, Cultured; D018925:Chemokines / Q000235:genetics / Q000276:immunology*; D002633:Chemotaxis; D006801:Humans; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D013290:Streptococcal Infections / Q000276:immunology* / Q000382:microbiology; D013292:Streptococcus agalactiae / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
chemical_list: D001426:Bacterial Proteins; D018925:Chemokines; D012697:Serine Endopeptidases