Identification of Conserved Domains in Salmonella muenchen Flagellin That Are Essential for Its Ability to Activate TLR5 and to Induce an Inflammatory Response in Vitro

Kanneganti G K Murthy, Amitabha Deb, Sunali Goonesekera, Csaba Szabo, Andrew L. Salzman

Research output: Contribution to journalArticle

89 Citations (Scopus)

Abstract

The bacterial surface protein flagellin is widely distributed and well conserved among distant bacterial species. We and other investigators have reported recently that purified flagellin from Salmonella dublin or recombinant flagellin of Salmonella muenchen origin binds to the eukaryotic toll receptor TLR5 and activates the nuclear translocation of NF-κB and mitogen-activated protein kinase, resulting in the release of a host of pro-inflammatory mediators in vitro and in vivo. The amino acid sequence alignment of flagellins from various Gram-negative bacteria shows that the C and N termini are well conserved. It is possible that sequences within the N and C termini or both may regulate the pro-inflammatory activity of flagellin. Here we set out to map more precisely the regions in both termini that are required for TLR5 activation and pro-inflammatory signaling. Systematic deletion of amino acids from either terminus progressively reduced eukaryotic pro-inflammatory activation. However, deletion of amino acids 95-108 (motif N) in the N terminus and 441-449 (motif C) in the C terminus abolished pro-inflammatory activity completely. Site-directed mutagenesis analysis provided further evidence for the importance of motifs N and C. We also present evidence for the functional role of motifs N and C with the TLR5 receptor using a reporter assay system. Taken together, our results demonstrate that the pro-inflammatory activity of flagellin results from the interaction of motif N with the TLR5 receptor on the cell surface.

Original languageEnglish (US)
Pages (from-to)5667-5675
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number7
DOIs
StatePublished - Feb 13 2004
Externally publishedYes

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Flagellin
Salmonella
Toll-Like Receptor 5
Amino Acids
Chemical activation
Mutagenesis
Bacterial Proteins
Sequence Alignment
Site-Directed Mutagenesis
Mitogen-Activated Protein Kinases
Gram-Negative Bacteria
In Vitro Techniques
Amino Acid Sequence
Assays
Bacteria
Membrane Proteins
Research Personnel

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification of Conserved Domains in Salmonella muenchen Flagellin That Are Essential for Its Ability to Activate TLR5 and to Induce an Inflammatory Response in Vitro. / Murthy, Kanneganti G K; Deb, Amitabha; Goonesekera, Sunali; Szabo, Csaba; Salzman, Andrew L.

In: Journal of Biological Chemistry, Vol. 279, No. 7, 13.02.2004, p. 5667-5675.

Research output: Contribution to journalArticle

Murthy, Kanneganti G K ; Deb, Amitabha ; Goonesekera, Sunali ; Szabo, Csaba ; Salzman, Andrew L. / Identification of Conserved Domains in Salmonella muenchen Flagellin That Are Essential for Its Ability to Activate TLR5 and to Induce an Inflammatory Response in Vitro. In: Journal of Biological Chemistry. 2004 ; Vol. 279, No. 7. pp. 5667-5675.
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