Salmonella enterica serotype Typhimurium ShdA is an outer membrane fibronectin-binding protein that is expressed in the intestine

Robert A. Kingsley, Renato L. Santos, A. Marijke Keestra, L. Garry Adams, Andreas J Baumler

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

The shdA gene is the only determinant known to be required for persistence of Salmonella enterica serotype Typhimurium (S. Typhimurium) in the murine caecum and for efficient and prolonged shedding of the organism with the faeces. To study the biological activity of the ShdA protein, we examined its expression and binding activity. ShdA was not detected with anti-ShdA antiserum in S. Typhimurium strain ATCC14028 grown in vitro, suggesting that this protein is not expressed under standard conditions of bacterial cultivation in the laboratory. However, in mice infected with S. Typhimurium, an immunofluorescence signal detected with anti-ShdA antiserum co-localized with that generated by anti-O4 antiserum in thin sections from the caecum. Expression of the cloned shdA gene from the T7 promoter in vitro resulted in detection of ShdA in the outer membrane of S. Typhimurium and in binding of fibronectin to the bacterial surface. Binding of purified glutathione-S-transferase (GST)-ShdA fusion protein to fibronectin was dose dependent and could be partially inhibited by preincubation with antifibronectin antibodies. GST-ShdA bound to connective tissue and the basement membrane in thin sections from the murine caecum in situ. A similar labelling pattern was produced when thin sections of the murine caecum were stained with antifibronectin antiserum. Collectively, these data demonstrate that ShdA is a surface-localized, fibronectin-binding protein whose expression is induced in vivo in the murine caecum, a tissue in which a cognate receptor of this outer membrane protein is expressed.

Original languageEnglish (US)
Pages (from-to)895-905
Number of pages11
JournalMolecular Microbiology
Volume43
Issue number4
DOIs
StatePublished - 2002
Externally publishedYes

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Salmonella enterica
Fibronectins
Intestines
Immune Sera
Carrier Proteins
Membranes
Glutathione Transferase
Proteins
Basement Membrane
Feces
Connective Tissue
Genes
Fluorescent Antibody Technique
Membrane Proteins
Serogroup
Antibodies
In Vitro Techniques

ASJC Scopus subject areas

  • Molecular Biology
  • Microbiology

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Salmonella enterica serotype Typhimurium ShdA is an outer membrane fibronectin-binding protein that is expressed in the intestine. / Kingsley, Robert A.; Santos, Renato L.; Keestra, A. Marijke; Adams, L. Garry; Baumler, Andreas J.

In: Molecular Microbiology, Vol. 43, No. 4, 2002, p. 895-905.

Research output: Contribution to journalArticle

Kingsley, Robert A. ; Santos, Renato L. ; Keestra, A. Marijke ; Adams, L. Garry ; Baumler, Andreas J. / Salmonella enterica serotype Typhimurium ShdA is an outer membrane fibronectin-binding protein that is expressed in the intestine. In: Molecular Microbiology. 2002 ; Vol. 43, No. 4. pp. 895-905.
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