Identification of a Porphyromonas gingivalis novel protein sov required for the secretion of gingipains.
Gingipains are extracellular proteases important for the virulence of Porphyromonas gingivalis; however, the mechanism for the secretion of gingipains is poorly understood. In this report, we found that insertion mutants for PG0809 (83K1 and 83K2) were defective in black pigmentation and hemolysis. We cloned and sequenced PG0809 and found that PG0809 contains two additional nucleotides that are not deposited in the W83 genome database. The revised sequence reveals an in-frame fusion of PG0810 and PG0809 and is designated the sov gene. We constructed a sov deletion mutant (83K3) and showed that 83K3 was defective in the activities of black pigmentation, hemolysis, and hemagglutination. Furthermore, in 83K3, the activities of gingipains were severely reduced whereas those of other secreted proteases DPPIV, DPP-7, and PtpA were not affected. Immunoblot analysis using anti-RgpB antiserum showed that Arg-gingipains were poorly secreted in an outer membrane or into an extracellular portion but accumulated within the cells of 83K3, suggesting the secretion of gingipains is defected in 83K3. Taken together, our findings indicated that Sov is a novel protein required for the secretion of gingipains and suggested that the secretion system for gingipains is different from the conserved secretion systems.
Saiki
Keitarou
K
Department of Microbiology, Nippon Dental University School of Life Dentistry at Tokyo, Chiyoda-ku, Tokyo 102-8159, Japan. keisaiki@tky.ndu.ac.jp
Konishi
Kiyoshi
K
D018829:Adhesins, Bacterial / Q000502:physiology*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000502:physiology*; D001483:Base Sequence; D003546:Cysteine Endopeptidases / Q000378:metabolism / Q000502:physiology*; D004269:DNA, Bacterial / Q000235:genetics; D004912:Erythrocytes / Q000502:physiology; D000080867:Gingipain Cysteine Endopeptidases; D006386:Hemagglutination Tests; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D016966:Porphyromonas gingivalis / Q000235:genetics / Q000502:physiology*; D040901:Proteomics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D000080867:Gingipain Cysteine Endopeptidases; D011994:Recombinant Proteins; D003546:Cysteine Endopeptidases