title: Porphyromonas gingivalis outer membrane vesicles exclusively contain outer membrane and periplasmic proteins and carry a cargo enriched with virulence factors.
abstract: Porphyromonas gingivalis, a keystone pathogen associated with chronic periodontitis, produces outer membrane vesicles (OMVs) that carry a cargo of virulence factors. In this study, the proteome of OMVs was determined by LC-MS/MS analyses of SDS-PAGE fractions, and a total of 151 OMV proteins were identified, with all but one likely to have originated from either the outer membrane or periplasm. Of these, 30 exhibited a C-terminal secretion signal known as the CTD that localizes them to the cell/vesicle surface, 79 and 27 were localized to the vesicle membrane and lumen respectively while 15 were of uncertain location. All of the CTD proteins along with other virulence factors were found to be considerably enriched in the OMVs, while proteins exhibiting the OmpA peptidoglycan-binding motif and TonB-dependent receptors were preferentially retained on the outer membrane of the cell. Cryo-transmission electron microscopy analysis revealed that an electron dense surface layer known to comprise CTD proteins accounted for a large proportion of the OMVs' volume providing an explanation for the enrichment of CTD proteins. Together the results show that P. gingivalis is able to specifically concentrate and release a large number of its virulence factors into the environment in the form of OMVs.
authors.affiliation: Oral Health CRC, Melbourne Dental School, Bio21 Institute, The University of Melbourne , 720 Swanston Street, Victoria, 3010, Australia.
authors.forename: Paul D
authors.initials: PD
authors.lastname: Veith
authors.forename: Yu-Yen
authors.initials: YY
authors.lastname: Chen
authors.forename: Dhana G
authors.initials: DG
authors.lastname: Gorasia
authors.forename: Dina
authors.initials: D
authors.lastname: Chen
authors.forename: Michelle D
authors.initials: MD
authors.lastname: Glew
authors.forename: Neil M
authors.initials: NM
authors.lastname: O'Brien-Simpson
authors.forename: Jessica D
authors.initials: JD
authors.lastname: Cecil
authors.forename: James A
authors.initials: JA
authors.lastname: Holden
authors.forename: Eric C
authors.initials: EC
authors.lastname: Reynolds
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002853:Chromatography, Liquid; D020285:Cryoelectron Microscopy; D004591:Electrophoresis, Polyacrylamide Gel; D046529:Microscopy, Electron, Transmission; D019897:Periplasm / Q000378:metabolism / Q000648:ultrastructure; D033903:Periplasmic Proteins / Q000378:metabolism*; D016966:Porphyromonas gingivalis / Q000378:metabolism* / Q000472:pathogenicity / Q000648:ultrastructure; D020543:Proteome / Q000378:metabolism; D040901:Proteomics / Q000379:methods; D015398:Signal Transduction; D053719:Tandem Mass Spectrometry; D022161:Transport Vesicles / Q000378:metabolism / Q000648:ultrastructure; D014774:Virulence; D037521:Virulence Factors / Q000378:metabolism*
chemical_list: D001425:Bacterial Outer Membrane Proteins; D033903:Periplasmic Proteins; D020543:Proteome; D037521:Virulence Factors