Immunological comparison of major outer membrane proteins from different strains of Escherichia coli

Pages, Catherine; Princé, P.; Pagès, Jean‐Marie

doi:10.1016/0769-2609(87)90057-3

Cited by 13 publications

(20 citation statements)

References 29 publications

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“…An extraction procedure which results in complete solubilization of the OmpF porin at low temperature was used (see Materials and methods). Immunoprecipitations were carried out with both a polyclonal antiserum which recognized all forms of OmpF (Pages et al, 1987), and MoF3, which detected only assembled trimers. The maximum of material obtained with the polyclonal antibodies was reached after correspond to films which were overexposed to obtain the same intensity of total immunoprecipitated OmpF (lanes 0-7) as for the control cells.…”

Section: Resultsmentioning

confidence: 99%

“…The ability to extract intact trimers and unassembled monomers has been used to examine the assembly of porins into the outer membrane (Baker et al, 1987), and an immunological approach to this problem has been successfully applied to the LamB protein (Vos-Scheperkeuter and Witholt, 1984;Stader and Silhavy, 1988). We recently described an immunological analysis of the various forms of OmpF in E. coli B (Pages et al, .1987;Pages and Bolla, 1988). This strain is particularly convenient because it contains only the OmpF porin, and we have obtained monoclonal antibodies specifically directed against regions of the trimer exposed at the surface of intact cells .…”

Section: Introductionmentioning

confidence: 99%

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The assembly of the major outer membrane protein OmpF of Escherichia coli depends on lipid synthesis.

1988

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Cerulenin, a drug which specifically blocks lipid synthesis, prevented both the trimerization of OmpF monomers and their assembly into the outer membrane of Escherichia coli B cells. A monoclonal antibody directed against a surface‐exposed epitope of the trimer was used to probe the assembly of OmpF in the presence or absence of the drug. An inhibition level of 80% was reached 16 min after the addition of cerulenin. The accumulated monomeric form could not be assembled even after lipid synthesis was restored. Instead, it was slowly degraded. It was further shown that the inhibition of assembly resulted in a rapid inhibition of OmpF synthesis. These data demonstrate that there is a direct relationship between the synthesis of lipid (most likely lipopolysaccharide) and the correct export of OmpF. This coupling is required to promote the trimerization of the porin monomer and its assembly into the outer membrane.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

The assembly of the major outer membrane protein OmpF of Escherichia coli depends on lipid synthesis.

1988

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“…Furthermore, to broaden the search for antigens, we included the LEE-negative O113:H11 serotype, which has also been associated with HUS (37,38). Prior studies have reported that exposure to enterobacterial OMPs stimulates a humoral immune response in infected patients (39)(40)(41), therefore making them candidates suitable for use in vaccines against Gram-negative pathogens (42)(43)(44)(45).…”

Section: Discussionmentioning

confidence: 99%

Immunoproteomic Analysis To Identify Shiga Toxin-Producing Escherichia coli Outer Membrane Proteins Expressed during Human Infection

et al. 2014

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b Shiga-toxin producing Escherichia coli (STEC) is the etiologic agent of acute diarrhea, dysentery, and hemolytic-uremic syndrome (HUS). There is no approved vaccine for STEC infection in humans, and antibiotic use is contraindicated, as it promotes Shiga toxin production. In order to identify STEC-associated antigens and immunogenic proteins, outer membrane proteins (OMPs) were extracted from STEC O26:H11, O103, O113:H21, and O157:H7 strains, and commensal E. coli strain HS was used as a control. SDS-PAGE, two-dimensional-PAGE analysis, Western blot assays using sera from pediatric HUS patients and controls, and matrix-assisted laser desorption ionization-tandem time of flight analyses were used to identify 12 immunogenic OMPs, some of which were not reactive with control sera. Importantly, seven of these proteins have not been previously reported to be immunogenic in STEC strains. Among these seven proteins, OmpT and Cah displayed IgG and IgA reactivity with sera from HUS patients. Genes encoding these two proteins were present in a majority of STEC strains. Knowledge of the antigens produced during infection of the host and the immune response to those antigens will be important for future vaccine development.

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“…Furthermore, some of these OMPs are target receptors for bacteriophages and colicins. Finally, due to their location as interfaces between the cell and the environment, OMPs are important candidate antigens for developing strategies aimed at providing protection against bacterial pathogens [7–10].…”

mentioning

confidence: 99%

Proteomic analysis of the Escherichia coli outer membrane

Molloy

Herbert²,

Slade

et al. 2000

European Journal of Biochemistry

441

396

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Outer membrane proteins (OMPs) of Gram-negative bacteria are key molecules that interface the cell with the environment. Traditional biochemical and genetic approaches have yielded a wealth of knowledge relating to the function of OMPs. Nonetheless, with the completion of the Escherichia coli genome sequencing project there is the opportunity to further expand our understanding of the organization, expression and function of the OMPs in this Gram-negative bacterium. In this report we describe a proteomic approach which provides a platform for parallel analysis of OMPs. We propose a rapid method for isolation of bacterial OMPs using carbonate incubation, purification and protein array by two-dimensional electrophoresis, followed by protein identification using mass spectrometry. Applying this method to examine E. coli K-12 cells grown in minimal media we identified 21 out of 26 (80%) of the predicted integral OMPs that are annotated in SWISS-PROT release 37 and predicted to separate within the range of pH 4±7 and molecular mass 10±80 kDa. Five outer membrane lipoproteins were also identified and only minor contamination by nonmembrane proteins was observed. Importantly, this research readily demonstrates that integral OMPs, commonly missing from 2D gel maps, are amenable to separation by two-dimensional electrophoresis. Two of the identified OMPs (YbiL, YeaF) were previously known only from their ORFs, and their identification confirms the cognate genes are transcribed and translated. Furthermore, we show that like the E. coli iron receptors FhuE and FhuA, the expression of YbiL is markedly increased by iron limitation, suggesting a putative role for this protein in iron transport. In an additional demonstration we show the value of parallel protein analysis to document changes in E. coli OMP expression as influenced by culture temperature.

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Immunological comparison of major outer membrane proteins from different strains of Escherichia coli

Cited by 13 publications

References 29 publications

The assembly of the major outer membrane protein OmpF of Escherichia coli depends on lipid synthesis.

The assembly of the major outer membrane protein OmpF of Escherichia coli depends on lipid synthesis.

Immunoproteomic Analysis To Identify Shiga Toxin-Producing Escherichia coli Outer Membrane Proteins Expressed during Human Infection

Proteomic analysis of the Escherichia coli outer membrane

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