Binding specificity of Salmonella plasmid-encoded fimbriae assessed by glycomics

Daniela Chessa, Caleb W. Dorsey, Maria Winter, Andreas J Baumler

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

The Salmonella enterica serotype Typhimurium (S. Typhimurium) genome encodes 12 intestinal colonization factors of the chaperone/usher fimbrial assembly class; however, the binding specificity is known for only one of these adhesins, known as type 1 fimbriae. Here we explored the utility of glycomics to determine the carbohydrate binding specificity of plasmid-encoded fimbriae from S. Typhimurium. A cosmid carrying the pef operon was introduced into Escherichia coli and expression of fimbrial filaments composed of PefA confirmed by flow cytometry and immune-electron microscopy. Plasmid-encoded fimbriae were purified from the surface of E. coli, and the resulting preparation was shown to contain PefA as the sole major protein component. The binding of purified plasmid-encoded fimbriae to a glycanarray suggested that this adhesin specifically binds the trisaccharide Galβ1-4(Fucα1-3)GlcNAc, also known as the Lewis X (Lex) blood group antigen. Results from the glycanarray were validated by enzyme-linked immunosorbent assay (ELISA) in which plasmid-encoded fimbriae bound Lex-coated wells in a concentration-dependent manner. The binding of plasmid-encoded fimbriae to Lex-coated wells could be inhibited by co-incubation with soluble Lex antigen. Our results establish glycomic analysis as a promising new approach for determining the carbohydrate binding specificity of bacterial adhesins.

Original languageEnglish (US)
Pages (from-to)8118-8124
Number of pages7
JournalJournal of Biological Chemistry
Volume283
Issue number13
DOIs
StatePublished - Mar 28 2008

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Glycomics
Salmonella
Plasmids
CD15 Antigens
Escherichia coli
Bacterial Adhesins
Carbohydrates
Trisaccharides
Immunosorbents
Cosmids
Salmonella enterica
Flow cytometry
Operon
Blood Group Antigens
Electron microscopy
Assays
Electron Microscopy
Flow Cytometry
Genes
Enzyme-Linked Immunosorbent Assay

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Binding specificity of Salmonella plasmid-encoded fimbriae assessed by glycomics. / Chessa, Daniela; Dorsey, Caleb W.; Winter, Maria; Baumler, Andreas J.

In: Journal of Biological Chemistry, Vol. 283, No. 13, 28.03.2008, p. 8118-8124.

Research output: Contribution to journalArticle

Chessa, Daniela ; Dorsey, Caleb W. ; Winter, Maria ; Baumler, Andreas J. / Binding specificity of Salmonella plasmid-encoded fimbriae assessed by glycomics. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 13. pp. 8118-8124.
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