Proteomic analysis of outer membrane vesicles derived from <i>Pseudomonas aeruginosa</i>

Choi, Dong-Sic; Kim, Dae-Kyum; Choi, Sang Joon; Lee, Jae‐Wook; Choi, Jun-Pyo; Rho, Sangchul; Park, Sang‐Hyun; Kim, Eui Chong; Hwang, Daehee; Gho, Yong Song

doi:10.1002/pmic.201000212

Cited by 211 publications

(205 citation statements)

References 41 publications

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“…The current understanding of FliC function localizes the protein in the flagellum outside the cell, while the ATP- dependent DnaK chaperone is considered to be cytoplasmic. However, E. coli FliC and several other FliC proteins, including P. aeruginosa FliC, were detected in the inner membrane fraction and the periplasm during various proteomic analyses (38)(39)(40)(41)(42)(43). In multiple approaches, the FliC protein was detected together with NirS (41) or even with NirS and DnaK (43).…”

Section: Discussionmentioning

confidence: 99%

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

et al. 2015

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Pseudomonas aeruginosa is a ubiquitously occurring environmental bacterium and opportunistic pathogen responsible for various acute and chronic infections. Obviously, anaerobic energy generation via denitrification contributes to its ecological success. To investigate the structural basis for the interconnection of the denitrification machinery to other essential cellular processes, we have sought to identify the protein interaction partners of the denitrification enzyme nitrite reductase NirS in the periplasm. We employed NirS as an affinity-purifiable bait to identify interacting proteins in vivo. Results obtained revealed that both the flagellar structural protein FliC and the protein chaperone DnaK form a complex with NirS in the periplasm. The interacting domains of NirS and FliC were tentatively identified. The NirS-interacting stretch of amino acids lies within its cytochrome c domain. Motility assays and ultrastructure analyses revealed that a nirS mutant was defective in the formation of flagella and correspondingly in swimming motility. In contrast, the fliC mutant revealed an intact denitrification pathway. However, deletion of the nirF gene, coding for a heme d 1 biosynthetic enzyme, which leads to catalytically inactive NirS, did not abolish swimming ability. This pointed to a structural function for the NirS protein. FliC and NirS were found colocalized with DnaK at the cell surface of P. aeruginosa. A function of the detected periplasmic NirS-DnaK-FliC complex in flagellum formation and motility was concluded and discussed. IMPORTANCEPhysiological functions in Gram-negative bacteria are connected with the cellular compartment of the periplasm and its membranes. Central enzymatic steps of anaerobic energy generation and the motility mediated by flagellar activity use these cellular structures in addition to multiple other processes. Almost nothing is known about the protein network functionally connecting these processes in the periplasm. Here, we demonstrate the existence of a ternary complex consisting of the denitrifying enzyme NirS, the chaperone DnaK, and the flagellar protein FliC in the periplasm of the pathogenic bacterium P. aeruginosa. The dependence of flagellum formation and motility on the presence of an intact NirS was shown, structurally connecting both cellular processes, which are important for biofilm formation and pathogenicity of the bacterium. P seudomonas aeruginosa is a metabolically versatile bacterium inhabiting multiple environmental niches (1). It is known for its highly efficient growth in the absence of oxygen. Fast anaerobic growth is mediated via the utilization of different N-oxides as electron acceptors in the respiratory chains of denitrification (2). Anaerobic respiratory growth via denitrification also sustains biofilm formation on environmental surfaces, in the mucus in the lung of cystic fibrosis patients, on the epithelium of the urinary tract infected individuals, and in burn wounds (3).The periplasm, the cellular compartment bounded by the cytoplasmi...

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Section: Discussionmentioning

confidence: 99%

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

et al. 2015

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“…Similar to the outer membrane, outer membrane vesicles (OMVs) possess an outer leaflet of lipopolysaccharide (LPS) and an inner leaflet of phospholipid (11)(12)(13)(14)(15). OMVs also contain outer membrane proteins and entrap periplasmic components as they are released (16)(17)(18). OMVs have been found to be associated with Gram-negative bacteria growing planktonically and in surface-attached biofilm communities as well as natural environments (11,(19)(20)(21).…”

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confidence: 99%

Role of Pseudomonas aeruginosa Peptidoglycan-Associated Outer Membrane Proteins in Vesicle Formation

et al. 2013

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Gram-negative bacteria produce outer membrane vesicles (OMVs) that package and deliver proteins, small molecules, and DNA to prokaryotic and eukaryotic cells. The molecular details of OMV biogenesis have not been fully elucidated, but peptidoglycanassociated outer membrane proteins that tether the outer membrane to the underlying peptidoglycan have been shown to be critical for OMV formation in multiple Enterobacteriaceae. In this study, we demonstrate that the peptidoglycan-associated outer membrane proteins OprF and OprI, but not OprL, impact production of OMVs by the opportunistic pathogen Pseudomonas aeruginosa. Interestingly, OprF does not appear to be important for tethering the outer membrane to peptidoglycan but instead impacts OMV formation through modulation of the levels of the Pseudomonas quinolone signal (PQS), a quorum signal previously shown by our laboratory to be critical for OMV formation. Thus, the mechanism by which OprF impacts OMV formation is distinct from that for other peptidoglycan-associated outer membrane proteins, including OprI.

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“…Global qualitative proteomic profiling has been conducted for OMVs of, e.g., Pseudomonas aeruginosa, Escherichia coli, Neisseria meningitidis, Myxococcus xanthus, and Acinetobacter baumannii primarily addressing the qualitative protein composition (8)(9)(10)(11)(12)(13). Reports addressing the selective sorting of distinct proteins into OMVs are rare (14).…”

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confidence: 99%

Comparative Proteome Analysis of Spontaneous Outer Membrane Vesicles and Purified Outer Membranes of Neisseria meningitidis

et al. 2013

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Proteomic analysis of outer membrane vesicles derived from Pseudomonas aeruginosa

Cited by 211 publications

References 41 publications

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

A Periplasmic Complex of the Nitrite Reductase NirS, the Chaperone DnaK, and the Flagellum Protein FliC Is Essential for Flagellum Assembly and Motility in Pseudomonas aeruginosa

Role of Pseudomonas aeruginosa Peptidoglycan-Associated Outer Membrane Proteins in Vesicle Formation

Comparative Proteome Analysis of Spontaneous Outer Membrane Vesicles and Purified Outer Membranes of Neisseria meningitidis

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