Konkel ME, Garvis SG, Tipton SL, et al. Identification and molecular cloning of a gene encoding a fibronectin-binding protein (CadF) from Campylobacter jejuni

Department of Microbiology, Washington State University, Pullman 99164-4233, USA.
Molecular Microbiology (Impact Factor: 5.03). 07/1997; 24(5):953-63. DOI: 10.1046/j.1365-2958.1997.4031771.x
Source: PubMed

ABSTRACT Campylobacter jejuni, a Gram-negative bacterium, is a common cause of gastrointestinal disease. By analogy with other enteric pathogens such as Salmonella and Shigella, the ability of C. jejuni to bind to host cells is thought to be essential in the pathogenesis of enteritis. Scanning electron microscopy of infected INT407 cells suggested that C. jejuni bound to a component of the extracellular matrix. Binding assays using immobilized extracellular matrix proteins and soluble fibronectin showed specific and saturable binding of fibronectin to C. jejuni. Ligand immunoblot assays using 125I-labelled fibronectin revealed specific binding to an outer membrane protein with an apparent molecular mass of 37 kDa. A rabbit antiserum, raised against the gel-purified protein, reacted with a 37 kDa protein in all C. jejuni isolates (n = 15) as tested by immunoblot analysis. Antibodies present in convalescent serum from C. jejuni-infected individuals also recognized a 37 kDa protein. The gene encoding the immunoreactive 37kDa protein was cloned and sequenced. Sequencing of overlapping DNA fragments revealed an open reading frame (ORF) that encodes a protein of 326 amino acids with a calculated molecular mass of 36872Da. The deduced amino acid sequence of the ORF exhibited 52% similarity and 28% identity to the root adhesin protein from Pseudomonas fluorescens. Isogenic C. jejuni mutants which lack the 37 kDa outer membrane protein, which we have termed CadF, displayed significantly reduced binding to fibronectin. Biotinylated fibronectin bound to a protein with an apparent molecular mass of 37 kDa in the outer membrane protein extracts from wild-type C. jejuni as judged by ligand-binding blots. These results indicate that the binding of C. jejuni to fibronectin is mediated by the 37 kDa outer membrane protein which is conserved among C. jejuni isolates.

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Available from: Steve Garvis, Oct 13, 2014
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    • "Also, Sanchez et al. found that biofilm bacteria grown on abiotic surfaces adhered better to epithelial cells than planktonic, broth grown bacteria (Sanchez et al., 2011b). These two studies, supported by studies in other human pathogens (Konkel et al., 1997; Sulaeman et al., 2012), demonstrate a relationship between epithelial cell adherence and biofilm formation, however, the studies have not investigated the role of this relationship during pneumococcal colonization. "
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    • "The pathogenicity of C. jejuni depends mainly on its ability to adhere and invade the cells of the human intestine. One of the adhesion factors used by C. jejuni to attach and eventually invade mammalian cells is CadF, a binding protein for fibronectin, a component of the extracellular matrix (Konkel et al. 1997, 2005). When C. jejuni enters the VBNC state, in response to environmental stress, it loses culturability, exhibits enhanced stress resistance, delays mouse lethality and modifies cell shape and protein profile (Tholozan et al. 1999; Baffone et al. 2006; Zhang et al. 2009). "
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    • "One explanation could be that the native Sub3 from M. canis is a cell-wall associated protein present on the surface of arthroconidia, interfering in adherence through intimate mechanisms that have not yet been elucidated. This has been notably reported for several pathogenic bacteria, such as Campylobacter jejuni (Konkel et al., 1997), some strains of Escherichia coli (Spencer et al., 1998) and Mannheimia haemolytica (Dagmara and Czuprynski, 2009). Fig. 3. Adherence of arthroconidia from the M. canis SUB3 RNA-silenced strain to feline epidermis in the presence of recombinant Sub3. "
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