title: PG27 is a novel membrane protein essential for a Porphyromonas gingivalis protease secretion system.
abstract: Porphyromonas gingivalis secretes endopeptidase gingipains, which are important virulence factors of this bacterium. Gingipains are transported across the inner membrane via the Sec system, followed by transport across the outer membrane via an unidentified pathway. The latter transport step is suggested to be mediated via a novel protein secretion pathway. In the present study, we report a novel candidate as an essential factor for the latter transport step. The PG0027 gene of P. gingivalis W83 encodes novel protein PG27. In a PG0027 deletion mutant (83K10), the activities of Arg-gingipain and Lys-gingipain were severely reduced, while the activities of secreted exopeptidases DPPIV, DPP-7, and PTP-A were unaffected. Protein localization was investigated by cell-surface biotinylation, subcellular fractionation, and immunoblot analysis. In the wild-type W83, Arg-gingipains in membrane fraction were detected as cell surface proteins. In contrast, in 83K10, Arg-gingipains were trapped in the periplasm and hardly secreted into an extracellular milieu. PG27 was suggested to be exposed to the cell surface by a cell surface biotinylation experiment; however, PG27 was detected in both inner and outer membrane fractions by subcellular fractionation experiments. Taken together, we suggest that PG27 is a unique membrane protein essential for a novel secretion pathway.
authors.affiliation: Department of Microbiology, Nippon Dental University School of Life Dentistry at Tokyo, Tokyo, Japan.
authors.forename: Ikumi
authors.initials: I
authors.lastname: Ishiguro
authors.forename: Keitarou
authors.initials: K
authors.lastname: Saiki
authors.forename: Kiyoshi
authors.initials: K
authors.lastname: Konishi
mesh_terms: D018829:Adhesins, Bacterial / Q000378:metabolism*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D019921:Biotinylation; D002458:Cell Fractionation; D002462:Cell Membrane / Q000737:chemistry; D003470:Culture Media / Q000737:chemistry; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D017353:Gene Deletion; D000080867:Gingipain Cysteine Endopeptidases; D015151:Immunoblotting; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D019897:Periplasm / Q000737:chemistry; D016966:Porphyromonas gingivalis / Q000235:genetics / Q000378:metabolism*
chemical_list: D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D003470:Culture Media; D000080867:Gingipain Cysteine Endopeptidases; D026901:Membrane Transport Proteins; D003546:Cysteine Endopeptidases