Sequence Variation in the Outer Membrane Protein-Encoding Gene cmeC, Conferring Multidrug Resistance among Campylobacter jejuni and Campylobacter coli Strains Isolated from Different Hosts

Campylobacter jejuni is an important cause of human foodborne gastroenteritis; strategies to prevent infection are hampered by a poor understanding of the complex interactions between host and pathogen. Previous work showed that C. jejuni could bind human histo-blood group antigens (BgAgs) in vitro and that BgAgs could inhibit the binding of C. jejuni to human intestinal mucosa ex vivo. Here, the major flagella subunit protein (FlaA) and the major outer membrane protein (MOMP) were identified as BgAg-binding adhesins in C. jejuni NCTC11168 . Significantly, the MOMP was shown to be O- glycosylated at Thr 268 ; previously only flagellin proteins were known to be O- glycosylated in C. jejuni . Substitution of MOMP Thr 268 led to significantly reduced binding to BgAgs. The O- glycan moiety was characterized as Gal(β1–3)-GalNAc(β1–4)-GalNAc(β1–4)-GalNAcα1-Thr 268 ; modelling suggested that O- glycosylation has a notable effect on the conformation of MOMP and this modulates BgAg-binding capacity. Glycosylation of MOMP at Thr 268 promoted cell-to-cell binding, biofilm formation and adhesion to Caco-2 cells, and was required for the optimal colonization of chickens by C. jejuni , confirming the significance of this O- glycosylation in pathogenesis.

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Point mutations in the major outer membrane protein drive hypervirulence of a rapidly expanding clone of Campylobacter jejuni

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1605869113 ◽

2016 ◽

Vol 113 (38) ◽

pp. 10690-10695 ◽

Cited By ~ 35

Author(s):

Zuowei Wu ◽

Balamurugan Periaswamy ◽

Orhan Sahin ◽

Michael Yaeger ◽

Paul Plummer ◽

...

Keyword(s):

Sequence Analysis ◽

Membrane Protein ◽

Outer Membrane ◽

Campylobacter Jejuni ◽

Outer Membrane Protein ◽

Animal Health ◽

Major Outer Membrane Protein ◽

Whole Genome Sequence ◽

Bacterial Strains ◽

Zoonotic Pathogen

Infections due to clonal expansion of highly virulent bacterial strains are clear and present threats to human and animal health. Association of genetic changes with disease is now a routine, but identification of causative mutations that enable disease remains difficult. Campylobacter jejuni is an important zoonotic pathogen transmitted to humans mainly via the foodborne route. C. jejuni typically colonizes the gut, but a hypervirulent and rapidly expanding clone of C. jejuni recently emerged, which is able to translocate across the intestinal tract, causing systemic infection and abortion in pregnant animals. The genetic basis responsible for this hypervirulence is unknown. Here, we developed a strategy, termed “directed genome evolution,” by using hybridization between abortifacient and nonabortifacient strains followed by selection in an animal disease model and whole-genome sequence analysis. This strategy successfully identified SNPs in porA, encoding the major outer membrane protein, are responsible for the hypervirulence. Defined mutagenesis verified that these mutations were both necessary and sufficient for causing abortion. Furthermore, sequence analysis identified porA as the gene with the top genome-wide signal of adaptive evolution using Fu’s Fs, a population genetic metric for recent population size changes, which is consistent with the recent expansion of clone “sheep abortion.” These results identify a key virulence factor in Campylobacter and a potential target for the control of this zoonotic pathogen. Furthermore, this study provides general, unbiased experimental and computational approaches that are broadly applicable for efficient elucidation of disease-causing mutations in bacterial pathogens.

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Campylobacter jejuni major outer membrane protein and a 59-kDa protein are involved in binding to fibronectin and INT 407 cell membranes

FEMS Microbiology Letters ◽

10.1111/j.1574-6968.1997.tb12778.x ◽

2006 ◽

Vol 157 (2) ◽

pp. 233-238 ◽

Cited By ~ 78

Author(s):

Irmgard Moser ◽

W Schroeder ◽

Johann Salnikow

Keyword(s):

Membrane Protein ◽

Outer Membrane ◽

Campylobacter Jejuni ◽

Outer Membrane Protein ◽

Cell Membranes ◽

Major Outer Membrane Protein

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Characterization of cross-reacting serotypes of Campylobacter jejuni

Canadian Journal of Microbiology ◽

10.1139/m89-040 ◽

1989 ◽

Vol 35 (2) ◽

pp. 265-273 ◽

Cited By ~ 22

Author(s):

Martin A. Preston ◽

J. L. Penner

Keyword(s):

Membrane Protein ◽

Outer Membrane ◽

Campylobacter Jejuni ◽

Outer Membrane Protein ◽

Antigenic Determinants ◽

Protein Patterns ◽

Cross Reactions ◽

Antigenic Properties ◽

Dna Restriction ◽

Reference Strains

Some strains of Campylobacter jejuni react with more than one reference antiserum from the serotyping scheme based on heat-stable lipopolysaccharide antigens. To investigate the molecular basis of these cross-reactions, lipopolysaccharides from the reference strains for serotypes 4, 13, 16, 43, and 50 and isolates recovered during two different outbreaks of C. jejuni enteritis were analyzed by passive haemagglutination and sodium dodecyl sulphate – polyacrylamide gel electrophoresis coupled with silver staining or immunoblotting. The results showed that lipopolysaccharides from the reference strains and the isolates reacted with antisera prepared against heterologous strains in various combinations and that both silver-stainable, low Mr and non-silver-stainable, high Mr lipopolysaccharide components provided the antigenic determinants associated with the cross-reactions. There were strain-to-strain differences in the structural and antigenic properties of these macromolecules and shared antigenic determinants were not always provided by a common structure. Analysis of the silver-stained lipopolysaccharide profiles, outer membrane protein patterns, and chromosomal DNA restriction patterns indicated that strains with the same lipopolysaccharide profile could have the same outer membrane protein pattern and the same DNA restriction pattern. These results provided evidence for the presence of clones within this antigenic complex and implicated antigenic variation in some strains as the phenomenon responsible for the multiplicity of cross-reactions.Key words: Campylobacter, serotypes, cross-reaction.

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Production of a Monoclonal Antibody Specific for the Major Outer Membrane Protein of Campylobacter jejuni and Characterization of the Epitope

Applied and Environmental Microbiology ◽

10.1128/aem.01559-07 ◽

2007 ◽

Vol 74 (3) ◽

pp. 833-839 ◽

Cited By ~ 7

Author(s):

Hongliang Qian ◽

Ervinna Pang ◽

Qingyun Du ◽

Jason Chang ◽

Jin Dong ◽

...

Keyword(s):

Membrane Protein ◽

Outer Membrane ◽

Campylobacter Jejuni ◽

Outer Membrane Protein ◽

Protein Band ◽

Major Outer Membrane Protein ◽

Dot Blot ◽

Detection And Identification ◽

Dot Blot Assay

ABSTRACT Campylobacter species are important enteric pathogens causing disease in humans and animals. There is a lack of a good immunological test that can be used routinely to separate Campylobacter jejuni from other Campylobacter species. We produced monoclonal antibodies (MAbs) directed against the major outer membrane protein (MOMP) of C. jejuni using recombinant MOMP as the antigen. One MAb, designated MAb5C4 and of the immunoglobulin G1 isotype, was found to be potentially specific for C. jejuni. Dot blots demonstrated that MAb5C4 reacted with all 29 isolates of C. jejuni tested but did not react with 2 C. jejuni isolates, 26 other Campylobacter spp. isolates, and 19 non-Campylobacter isolates. Western blotting showed that MAb5C4 bound to a single protein band approximately 43 kDa in size, corresponding to the expected size of C. jejuni MOMP. The detection limit of MAb5C4 in a dot blot assay was determined to be about 5 × 103 bacteria. The epitope on the MOMP was mapped to a region six amino acids in length with the sequence 216GGQFNP221, which is 97% conserved among C. jejuni strains but divergent in other Campylobacter spp.; a GenBank search indicated that 95% of C. jejuni isolates will be able to be detected from non-Campylobacter spp. based on the highly specific and conserved region of the GGQFNP polypeptide. The epitope is predicted to be located in a region that is exposed to the periplasm. MAb5C4 is a potentially specific and sensitive MAb that can be used for the specific detection and identification of C. jejuni.

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