JlpA, a novel surface-exposed lipoprotein specific to Campylobacter jejuni, mediates adherence to host epithelial cells.
A 1116 bp open reading frame (ORF), designated jlpA, encoding a novel species-specific lipoprotein of Campylobacter jejuni TGH9011, was identified from recombinant plasmid pHIP-O. The jlpA gene encodes a polypeptide (JlpA) of 372 amino acid residues with a molecular mass of 42.3 kDa. JlpA contains a typical signal peptide and lipoprotein processing site at the N-terminus. The presence of a lipid moiety on the JlpA molecule was confirmed by the incorporation of [3H]-palmitic acid. Immunoblotting analysis of cell surface extracts prepared using glycine-acid buffer (pH 2.2) and proteinase K digestion of whole cells indicated that JlpA is a surface-exposed lipoprotein in C. jejuni. JlpA is loosely associated with the cell surface, as it is easily extracted from the C. jejuni outer membrane by detergents, such as sarcosyl and Triton X-100. JlpA is released to the culture medium, and its concentration increases in a time-dependent fashion. The adherence of both insertion and deletion mutants of jlpA to HEp-2 epithelial cells was reduced compared with that of parental C. jejuni TGH9011. Adherence of C. jejuni to HEp-2 cells was inhibited in a dose-dependent manner when the bacterium was preincubated with anti-GST-JlpA antibodies or when HEp-2 cells were preincubated with JlpA protein. A ligand-binding immunoblotting assay showed that JlpA binds to HEp-2 cells, which suggests that JlpA is C. jejuni adhesin.
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Departments of Medical Genetics and Microbiology, Laboratory Medicine and Pathobiology, and Pediatrics, University of Toronto, Ontario, Canada M5S 1A8.
Joe
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Lynett
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Hani
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Sherman
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Chan
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VL
D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism*; D001422:Bacterial Adhesion; D002169:Campylobacter Infections / Q000382:microbiology; D016123:Campylobacter jejuni / Q000235:genetics / Q000472:pathogenicity* / Q000502:physiology; D002460:Cell Line; D003470:Culture Media; D004847:Epithelial Cells / Q000382:microbiology*; D006801:Humans; D008074:Lipoproteins / Q000235:genetics / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013045:Species Specificity; D014774:Virulence
D018829:Adhesins, Bacterial; D003470:Culture Media; D008074:Lipoproteins; D011994:Recombinant Proteins