Antigenic Determinants of Murein Lipoprotein and Its Exposure at the Surface of Enterobacteriaceae

Braun, Volkmar; Bosch, V.; Klumpp, Ernst Rudolf; Neff, Ingrid; Mayer, Hubert; Schlecht, S

doi:10.1111/j.1432-1033.1976.tb10190.x

Cited by 54 publications

(24 citation statements)

References 35 publications

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“…(ii) The spectrum of crossreactivity observed for antigen 47 parallels that observed here ( Fig. 6B and C) and recorded in part elsewhere (5,31,33) (39,41) (Fig. 1).…”

Section: Downloaded Fromsupporting

confidence: 67%

Identification, immunochemical characterization, and purification of a major lipoprotein antigen associated with the inner (cytoplasmic) membrane of Escherichia coli

Doherty¹,

Yamada²,

Caffrey³

et al. 1986

J Bacteriol

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A major antigenic constituent of the inner membrane of Escherichia coli ML308-225 was identified as a 28.5-kilodalton lipoprotein containing covalently bound glycerol and palmitate. This lipoprotein corresponded to antigen 47 in the crossed immunoelectrophoresis profile of membrane vesicles (P. Owen and H. R. Kaback, Proc. Natl. Acad. Sci. USA 75:3148-3152, 1978) and to new lipoprotein 4 described for E. coli B by Ichihara et al. (S. Ichihara, H. Hussain, and S. Mizushima, J. Biol. Chem. 256:3125-3129, 1980). Experiments involving isopycnic centrifugation of spheroplast envelopes indicated that antigen 47 was enriched in cytoplasmic membrane subfractions of low density. The protein did not manifest an obvious association with peptidoglycan of the types displayed by the bound form of the Braun (Lpp) lipoprotein, the 21-kilodalton peptidoglycan-associated lipoprotein, or the ompFIC gene products. Antibodies specific for antigen 47 were used to demonstrate that the molecule was immunologically distinct from both the Braun lipoprotein and the peptidoglycan-associated lipoprotein of E. coli. Antigens of similar molecular mass to and cross-reacting with antigen 47 were present in the envelopes of eight type species of the Enterobacteriaceae. A protocol for the purification of antigen 47, based upon its solubility in a chloroform-methanol-water mixture, was developed.The covalent attachment of lipid to proteins was first suggested over 30 years ago by Folch and Lees (16). Since that time, an increasing number of membrane proteins of both eucaryotic and procaryotic origin have been reported to contain covalently bound lipid (12,23,29,47,49,52). Common features of most procaryotic lipoproteins studied to date appear to be an N-acyl diglyceride cysteine at the N terminus of the molecule (18,25,35) and a posttranslational modification event involving a signal peptidase which can be selectively inhibited by the antibiotic globomycin (22,23,60). In gram-positive bacteria, the covalent association of membrane-bound penicillinases with lipid is well documented (25,34,35,37,53). In Escherichia coli, the envelope is known to contain a number of lipoproteins (4,23,30). Of these, the most dominant and best characterized are associated with the outer membrane system and include the Braun (Lpp) lipoprotein, the peptidoglycan-associated lipoprotein (PAL), and the traT gene product (4,20,30,31,48). The presence of several other (minor) envelope-associated lipoproteins (termed new lipoproteins or NLPs) has also been documented (23), and there is evidence that some of these, viz., NLP3, NLP4, and possibly NLP6, may reside in the inner membrane of E. coli (23, 32).Reports emanating from this laboratory over the course of the last 10 years (for reviews, see references 39-41) have been concerned with an analysis of the numerous antigens associated with the envelope of E. coli (50, 54). Our understanding of the complexity, identity, and topography of these antigens, many of which are proteinaceous in nature, has been facilitated by the us...

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“…(ii) The spectrum of crossreactivity observed for antigen 47 parallels that observed here ( Fig. 6B and C) and recorded in part elsewhere (5,31,33) (39,41) (Fig. 1).…”

Section: Downloaded Fromsupporting

confidence: 67%

Identification, immunochemical characterization, and purification of a major lipoprotein antigen associated with the inner (cytoplasmic) membrane of Escherichia coli

Doherty¹,

Yamada²,

Caffrey³

et al. 1986

J Bacteriol

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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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“…OmpA, PAL, and MLP are structural outer membrane proteins (3,10,39,45,54,58,61) that are highly conserved among different enteric gram-negative bacteria (2,8,33,47). Proteins similar to each exist in nonenteric gramnegative bacteria (46,48,59).…”

Section: Discussionmentioning

confidence: 99%

Outer Membrane Protein A, Peptidoglycan-Associated Lipoprotein, and Murein Lipoprotein Are Released byEscherichia coliBacteria into Serum

Hellman

Loiselle²,

Tehan

et al. 2000

Infect Immun

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4Complexes containing lipopolysaccharide (LPS) and three outer membrane proteins (OMPs) are released by gram-negative bacteria incubated in human serum and into the circulation in an experimental model of sepsis. The same OMPs are bound by immunoglobulin G (IgG) in the cross-protective antiserum raised to Escherichia coli J5 (anti-J5 IgG). This study was performed to identify the three OMPs. The 35-kDa OMP was identified as outer membrane protein A (OmpA) by immunoblotting studies using OmpA-deficient bacteria and recombinant OmpA protein. The 18-kDa OMP was identified as peptidoglycan-associated lipoprotein (PAL) based on peptide sequences from the purified protein and immunoblotting studies using PAL-deficient bacteria. The 5-to 9-kDa OMP was identified as murein lipoprotein (MLP) based on immunoblotting studies using MLPdeficient bacteria. The studies identify the OMPs released into human serum and into the circulation in an experimental model of sepsis as OmpA, PAL, and MLP.Bacterial cell wall components released into the bloodstream are believed to be important in the pathogenesis of gram-negative sepsis. Although prior investigators have reported that bacteria release lipopolysaccharide (LPS) into serum (62, 63) and into the circulation (4,18,56,66), the full composition of released bacterial products has not been established. Very little is known about release of non-LPS gramnegative outer membrane components such as outer membrane proteins (OMPs) in sepsis. Fragments containing LPS, OmpA, and another faintly staining protein, of 17 kDa, were affinity purified from filtrates of human serum incubated with Salmonella enterica serovar Abortus equi bacteria using Ochain-specific anti-LPS immunoglobulin G (IgG) (20). Similarly, we have affinity purified complexes containing LPS and at least three OMPs, with estimated molecular masses of 35, 18, and 5 to 9 kDa, from filtrates of normal human serum incubated with Escherichia coli bacteria, using O-chain-specific anti-LPS IgG (29,30).Previous studies indicated that passive and active immunity directed to rough mutant bacteria such as S. enterica serovar Minnesota Re595 and E. coli J5 protect in experimental and clinical gram-negative sepsis (1,5,11,42,43,68). Protection has been attributed to antibodies directed to conserved core components of LPS (lipid A and core oligosaccharide). However, it has been difficult to prove that antisera to rough strains of bacteria contain cross-reactive anti-lipid A or anti-core oligosaccharide IgGs (15, 57), and the exact mechanism of protection remains unclear and controversial.We have demonstrated that IgG in antiserum raised to heatkilled E. coli J5 (J5 antiserum) binds to the same three gramnegative bacterial OMPs that are released into serum in the OMP-LPS complexes described above (30). OMP-LPS complexes are also released into the bloodstream of burned rats with E. coli O18K ϩ sepsis (29). In addition, at least one OMP, with an estimated molecular mass of 18 kDa, is released from bacteria separately from the OMP-LPS comple...

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Antigenic Determinants of Murein Lipoprotein and Its Exposure at the Surface of Enterobacteriaceae

Cited by 54 publications

References 35 publications

Identification, immunochemical characterization, and purification of a major lipoprotein antigen associated with the inner (cytoplasmic) membrane of Escherichia coli

Identification, immunochemical characterization, and purification of a major lipoprotein antigen associated with the inner (cytoplasmic) membrane of Escherichia coli

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

Outer Membrane Protein A, Peptidoglycan-Associated Lipoprotein, and Murein Lipoprotein Are Released byEscherichia coliBacteria into Serum

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