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  1. Article ; Online: Klebsiella pneumoniae O antigen loss alters the outer membrane protein composition and the selective packaging of proteins into secreted outer membrane vesicles.

    Cahill, Bethaney K / Seeley, Kent W / Gutel, Dedra / Ellis, Terri N

    Microbiological research

    2015  Volume 180, Page(s) 1–10

    Abstract: Klebsiella pneumoniae is a nosocomial pathogen which naturally secretes lipopolysaccharide (LPS) and cell envelope associated proteins into the environment through the production of outer membrane vesicles (OMVs). The loss of the LPS O antigen has been ... ...

    Abstract Klebsiella pneumoniae is a nosocomial pathogen which naturally secretes lipopolysaccharide (LPS) and cell envelope associated proteins into the environment through the production of outer membrane vesicles (OMVs). The loss of the LPS O antigen has been demonstrated in other bacterial species to significantly alter the composition of OMVs. Therefore, this study aimed to comprehensively analyze the impact of O antigen loss on the sub-proteomes of both the outer membrane and secreted OMVs from K. pneumoniae. As determined by LC-MS/MS, OMVs were highly enriched with outer membrane proteins involved in cell wall, membrane, and envelope biogenesis as compared to the source cellular outer membrane. Deletion of wbbO, the enzyme responsible for O antigen attachment to LPS, decreased but did not eliminate this enrichment effect. Additionally, loss of O antigen resulted in OMVs with increased numbers of proteins involved in post-translational modification, protein turnover, and chaperones as compared to secreted vesicles from the wild type. This alteration of OMV composition may be a compensatory mechanism to deal with envelope stress. This comprehensive analysis confirms the highly distinct protein composition of OMVs as compared to their source membrane, and provides evidence for a selective sorting mechanism that involves LPS polysaccharides. These data support the hypothesis that modifications to LPS alters both the mechanics of protein sorting and the contents of secreted OMVs and significantly impacts the protein composition of the outer membrane.
    MeSH term(s) Bacterial Outer Membrane Proteins/isolation & purification ; Bacterial Outer Membrane Proteins/metabolism ; Cell Membrane/metabolism ; Klebsiella pneumoniae/cytology ; Klebsiella pneumoniae/genetics ; Klebsiella pneumoniae/immunology ; Klebsiella pneumoniae/metabolism ; Molecular Chaperones/metabolism ; Mutation ; O Antigens/genetics ; O Antigens/metabolism ; Protein Processing, Post-Translational ; Protein Transport ; Proteome/genetics ; Proteome/secretion ; Tandem Mass Spectrometry ; Virulence Factors/metabolism
    Chemical Substances Bacterial Outer Membrane Proteins ; Molecular Chaperones ; O Antigens ; Proteome ; Virulence Factors
    Language English
    Publishing date 2015-11
    Publishing country Germany
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 1189614-0
    ISSN 1618-0623 ; 0944-5013
    ISSN (online) 1618-0623
    ISSN 0944-5013
    DOI 10.1016/j.micres.2015.06.012
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Klebsiella pneumoniae O antigen loss alters the outer membrane protein composition and the selective packaging of proteins into secreted outer membrane vesicles

    Cahill, Bethaney K / Dedra Gutel / Kent W. Seeley / Terri N. Ellis

    Microbiological research. 2015 Nov., v. 180

    2015  

    Abstract: Klebsiella pneumoniae is a nosocomial pathogen which naturally secretes lipopolysaccharide (LPS) and cell envelope associated proteins into the environment through the production of outer membrane vesicles (OMVs). The loss of the LPS O antigen has been ... ...

    Abstract Klebsiella pneumoniae is a nosocomial pathogen which naturally secretes lipopolysaccharide (LPS) and cell envelope associated proteins into the environment through the production of outer membrane vesicles (OMVs). The loss of the LPS O antigen has been demonstrated in other bacterial species to significantly alter the composition of OMVs. Therefore, this study aimed to comprehensively analyze the impact of O antigen loss on the sub-proteomes of both the outer membrane and secreted OMVs from K. pneumoniae. As determined by LC–MS/MS, OMVs were highly enriched with outer membrane proteins involved in cell wall, membrane, and envelope biogenesis as compared to the source cellular outer membrane. Deletion of wbbO, the enzyme responsible for O antigen attachment to LPS, decreased but did not eliminate this enrichment effect. Additionally, loss of O antigen resulted in OMVs with increased numbers of proteins involved in post-translational modification, protein turnover, and chaperones as compared to secreted vesicles from the wild type. This alteration of OMV composition may be a compensatory mechanism to deal with envelope stress. This comprehensive analysis confirms the highly distinct protein composition of OMVs as compared to their source membrane, and provides evidence for a selective sorting mechanism that involves LPS polysaccharides. These data support the hypothesis that modifications to LPS alters both the mechanics of protein sorting and the contents of secreted OMVs and significantly impacts the protein composition of the outer membrane.
    Keywords antigens ; biogenesis ; cell walls ; cross infection ; Klebsiella pneumoniae ; lipopolysaccharides ; mechanics ; outer membrane proteins ; packaging ; pathogens ; post-translational modification ; protein composition ; protein secretion ; protein transport
    Language English
    Dates of publication 2015-11
    Size p. 1-10.
    Publishing place Elsevier GmbH
    Document type Article
    ZDB-ID 1189614-0
    ISSN 1618-0623 ; 0944-5013
    ISSN (online) 1618-0623
    ISSN 0944-5013
    DOI 10.1016/j.micres.2015.06.012
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Porin Loss Impacts the Host Inflammatory Response to Outer Membrane Vesicles of Klebsiella pneumoniae.

    Turner, Kelli L / Cahill, Bethaney K / Dilello, Sarah K / Gutel, Dedra / Brunson, Debra N / Albertí, Sebastián / Ellis, Terri N

    Antimicrobial agents and chemotherapy

    2015  Volume 60, Issue 3, Page(s) 1360–1369

    Abstract: Antibiotic-resistant strains of Klebsiella pneumoniae often exhibit porin loss. In this study, we investigated how porin loss impacted the composition of secreted outer membrane vesicles as well as their ability to trigger proinflammatory cytokine ... ...

    Abstract Antibiotic-resistant strains of Klebsiella pneumoniae often exhibit porin loss. In this study, we investigated how porin loss impacted the composition of secreted outer membrane vesicles as well as their ability to trigger proinflammatory cytokine secretion by macrophages. We hypothesize that porin loss associated with antibiotic resistance will directly impact both the composition of outer membrane vesicles and their interactions with phagocytic cells. Using clonally related clinical isolates of extended-spectrum beta-lactamase (ESBL)-positive Klebsiella pneumoniae with different patterns of porin expression, we demonstrated that altered expression of OmpK35 and OmpK36 results in broad alterations to the protein profile of secreted vesicles. Additionally, the level of OmpA incorporation was elevated in strains lacking a single porin. Porin loss significantly impacted macrophage inflammatory responses to purified vesicles. Outer membrane vesicles lacking both OmpK35 and OmpK36 elicited significantly lower levels of proinflammatory cytokine secretion than vesicles from strains expressing one or both porins. These data demonstrate that antibiotic resistance-associated porin loss has a broad and significant effect on both the composition of outer membrane vesicles and their interactions with phagocytic cells, which may impact bacterial survival and inflammatory reactions in the host.
    MeSH term(s) Animals ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Cell Line ; Gene Expression Regulation, Bacterial ; Granulocyte-Macrophage Colony-Stimulating Factor/metabolism ; Host-Pathogen Interactions/physiology ; Inflammation/microbiology ; Klebsiella pneumoniae/growth & development ; Klebsiella pneumoniae/pathogenicity ; Macrophages/metabolism ; Macrophages/microbiology ; Macrophages/pathology ; Mice ; Porins/genetics ; Porins/metabolism ; Secretory Vesicles/genetics ; Secretory Vesicles/metabolism ; Tumor Necrosis Factor-alpha/metabolism
    Chemical Substances Bacterial Proteins ; OmpK35 porin, Klebsiella pneumoniae ; OmpK36 protein, Klebsiella pneumoniae ; Porins ; Tumor Necrosis Factor-alpha ; Granulocyte-Macrophage Colony-Stimulating Factor (83869-56-1)
    Language English
    Publishing date 2015-12-14
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 217602-6
    ISSN 1098-6596 ; 0066-4804
    ISSN (online) 1098-6596
    ISSN 0066-4804
    DOI 10.1128/AAC.01627-15
    Database MEDical Literature Analysis and Retrieval System OnLINE

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