title: Effect of group A streptococcal cysteine protease on invasion of epithelial cells.
abstract: Cysteine protease of group A streptococci (GAS) is considered an important virulence factor. However, its role in invasiveness of GAS has not been investigated. We demonstrated in this study that two strains of protease-producing GAS had the ability to invade A-549 human respiratory epithelial cells. Isogenic protease mutants were constructed by using integrational plasmids to disrupt the speB gene and confirmed by Southern hybridization and Western immunoblot analyses. No extracellular protease activity was produced by the mutants. The mutants had growth rates similar to those of the wild-type strains and produced normal levels of other extracellular proteins. When invading A-549 cells, the mutants had a two- to threefold decrease in activity compared to that of the wild-type strains. The invasion activity increased when the A-549 cells were incubated with purified cysteine protease and the mutant. However, blockage of the cysteine protease with a specific cysteine protease inhibitor, E-64, decreased the invasion activity of GAS. Intracellular growth of GAS was not found in A-549 cells. The presence or absence of protease activity did not affect the adhesive ability of GAS. These results suggested that streptococcal cysteine protease can enhance the invasion ability of GAS in human respiratory epithelial cells.
authors.affiliation: Department of Microbiology and Immunology, National Cheng Kung University Medical College, Tainan, Taiwan, Republic of China.
authors.forename: P J
authors.initials: PJ
authors.lastname: Tsai
authors.forename: C F
authors.initials: CF
authors.lastname: Kuo
authors.forename: K Y
authors.initials: KY
authors.lastname: Lin
authors.forename: Y S
authors.initials: YS
authors.lastname: Lin
authors.forename: H Y
authors.initials: HY
authors.lastname: Lei
authors.forename: F F
authors.initials: FF
authors.lastname: Chen
authors.forename: J R
authors.initials: JR
authors.lastname: Wang
authors.forename: J J
authors.initials: JJ
authors.lastname: Wu
mesh_terms: D001422:Bacterial Adhesion; D001426:Bacterial Proteins; D002460:Cell Line; D003546:Cysteine Endopeptidases / Q000235:genetics / Q000502:physiology*; D004847:Epithelial Cells / Q000382:microbiology; D006801:Humans; D011650:Pulmonary Alveoli / Q000382:microbiology; D013297:Streptococcus pyogenes / Q000201:enzymology* / Q000254:growth & development
chemical_list: D001426:Bacterial Proteins; D003546:Cysteine Endopeptidases; C042477:streptopain