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Three conserved MyD88-recruiting TLR residues exert different effects on the human TLR4 signaling pathway

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Abstract

Stimulation of Toll-like receptor (TLR) 4 leads to the activation of both MyD88-dependent and MyD88-independent pathways through the recruitment of adaptors TIRAP/MyD88 and TRIF/TRAM, respectively. However, the molecular basis of the TLR4 Toll/interleukin-1 receptor (TIR) domain in recruiting these downstream adaptors is still not entirely clear. Here, we identify three amino acid residues (714P in the BB loop, 696L in the αA helix and 721N in the αB sheet) conserved in all MyD88-recruited TLRs, but not the TLR3 TIR domain, as being critical for TLR4 responsiveness to LPS. These results were based on the substitution of each residue with a residue of the opposite type (hydrophilic/hydrophobic). However, the responsiveness of the TLR4 mutants to LPS was only partially decreased when each residue was replaced with a residue having the same hydrophilicity/hydrophobicity. This result is likely associated with an alteration in the BB-loop conformation of each TLR4 mutant and its ability to recruit the downstream adaptor TRAM. Thus, we identified three amino acids essential for TLR4 signaling, and their replacement with a residue of the same or opposite hydrophilicity/hydrophobicity greatly affected TLR4 signaling. This study furthers our understanding of the molecular mechanism by which the TLR4 TIR domain modulates TLR4 signaling and also provides new insight for the design of antisepsis therapy.

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Acknowledgments

This work is supported by the Natural Science Foundation of China for WX (81000747) and the Natural Science Foundation of Military for WX (CWS12J093). We are grateful to Professor Kensuke Miyake for providing us with plasmid pEF-BOS-TLR4 and pEF-BOS-MD-2 and Dr. Xin Du for pFLAG-CMV1-CD14.

Conflict of interest

The authors declare that there is no conflict of interest.

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Authors and Affiliations

  1. Department of Pathogenic Biology, Third Military Medical University, 30 Gaotanyan Zhengjie, Shapingba District, Chongqing, 400038, People’s Republic of China

    Yan Ding, Yuan Qiu, Zhangping Tan & Wenyue Xu

  2. Department of Microbiology, Third Military Medical University, Chongqing, 400038, People’s Republic of China

    Linyun Zou

  3. Department of Thoracic Surgery of Xinqiao Hospital, Third Military Medical University, Chongqing, 400037, People’s Republic of China

    Jigang Dai

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  1. Yan Ding
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  2. Yuan Qiu
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  3. Linyun Zou
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  4. Zhangping Tan
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  5. Jigang Dai
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  6. Wenyue Xu
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Correspondence to Jigang Dai or Wenyue Xu.

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Ding, Y., Qiu, Y., Zou, L. et al. Three conserved MyD88-recruiting TLR residues exert different effects on the human TLR4 signaling pathway. Immunol Res 62, 213–221 (2015). https://doi.org/10.1007/s12026-015-8652-2

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