Construction and Characterization of Arginine-specific Cysteine Proteinase (Arg-gingipain)-deficient Mutants of Porphyromonas gingivalis EVIDENCE FOR SIGNIFICANT CONTRIBUTION OF Arg-GINGIPAIN TO VIRULENCE

摘要: Arginine-specific cysteine proteinase (Arg-gingipain; formerly, argingipain) is one of the major extracellular proteinases produced by oral anaerobic bacterium Porphyromonas gingivalis. To determine whether Arg-gingipain important for periodontopathogenicity organism, Arg-gingipain-deficient mutants were constructed via gene disruption use suicide plasmid systems. First, Southern hybridization analyses suggested that two separate Arg-gingipain-encoding genes designated rgpA and rgpB existed on 12.5- 7.8-kilobase pair HindIII chromosomal fragments P. gingivalis ATCC33277, respectively. single mobilization a plasmid. Then, an double mutant was isolated electroporation with second No proteolytic activity observed in either cell extract or culture supernatant mutant. The chemiluminescence response polymorphonuclear leukocytes, which closely related to their bactericidal function, not inhibited mutant, while wild type parent showed significant inhibition response. result suggests responsible function leukocytes. In addition, mutations caused marked decrease hemagglutination gingivalis, indicating part hemagglutinin organism associated genes. These findings demonstrate makes contribution virulence