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Toll-like receptor 5 recognizes a conserved site on flagellin required for protofilament formation and bacterial motility

Nature Immunology volume 4, pages 1247–1253 (2003)Cite this article

A Corrigendum to this article was published on 01 April 2004

Abstract

Toll-like receptor 5 (TLR5) recognizes bacterial flagellin and activates host inflammatory responses. In this study, we examine the nature of the TLR5-flagellin interaction. With deletional, insertional and alanine-scanning mutagenesis, we precisely mapped the TLR5 recognition site on flagellin to a cluster of 13 amino acid residues that participate in intermolecular interactions within flagellar protofilaments and that are required for bacterial motility. The recognition site is buried in the flagellar filament, and monomeric flagellin, but not the filamentous molecule, stimulated TLR5. Finally, flagellin coprecipitated with TLR5, indicating close physical interaction between the molecules. These studies demonstrate the exquisite ability of the innate immune system to precisely target a conserved site on flagellin that is essential for bacterial motility.

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Figure 1: D1 domain N- and C-terminal deletions abrogates TLR5 recognition.
Figure 2: TLR5 recognizes discrete site in the D1 domain.
Figure 3: Flagellin alanine point mutations reduce TLR5 recognition.
Figure 4: TLR5 recognizes monomeric and not filamentous flagellin.
Figure 5: Biotinylated flagellin specifically associates with TLR5.

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Acknowledgements

We thank D. Underhill, A. Ozinsky and members of the Aderem laboratory for their comments and suggestions. This work was supported by National Institutes of Health grants K08AI01751 (K.D.S.); R01AI032972, R37AI025032 and R01AI52286 (A.A.); and R01AI047242 (B.T.C.).

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  1. Fumitaka Hayashi

    Present address: Center for Immunology and Inflammatory Diseases and Division of Rheumatology, Allergy and Immunology, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts, 02129–2020, USA

Authors and Affiliations

  1. Institute for Systems Biology, 1441 North 34th Street, Seattle, 98103, Washington, USA

    Kelly D Smith, Erica Andersen-Nissen, Fumitaka Hayashi, Katie Strobe & Alan Aderem

  2. Department of Pathology, the University of Washington, 1959 NE Pacific Street, Seattle, 98195, Washington, USA

    Kelly D Smith

  3. Department of Immunology, the University of Washington, 1959 NE Pacific Street, Seattle, 98195, Washington, USA

    Erica Andersen-Nissen & Fumitaka Hayashi

  4. Department of Microbiology, the University of Washington, 1959 NE Pacific Street, Seattle, 98195, Washington, USA

    Molly A Bergman, Sara L Rassoulian Barrett & Brad T Cookson

  5. Department of Laboratory Medicine, the University of Washington, 1959 NE Pacific Street, Seattle, 98195, Washington, USA

    Brad T Cookson

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Smith, K., Andersen-Nissen, E., Hayashi, F. et al. Toll-like receptor 5 recognizes a conserved site on flagellin required for protofilament formation and bacterial motility. Nat Immunol 4, 1247–1253 (2003). https://doi.org/10.1038/ni1011

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