Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A

Natsuko Yamakawa, Umeharu Ohto, Sachiko Akashi-Takamura, Koichiro Takahashi, Shin Ichiroh Saitoh, Natsuko Tanimura, Takayoshi Suganami, Yoshihiro Ogawa, Takuma Shibata, Toshiyuki Shimizu, Kensuke Miyake

Research output: Contribution to journalArticle

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Abstract

A cell surface heterodimer Toll-like receptor 4 (TLR4)/MD-2 senses lipopolysaccharide (LPS), a principal membrane component of Gram-negative bacteria. LPS binds to MD-2 and induces dimerization of TLR4/MD-2. Dimerized TLR4 activates downstream signaling. TLR4 polymorphism replacing Asp299 with Gly and Thr399 with Ile (D299G/T399I) causes LPS hyporesponsiveness, and is associated with a variety of infectious and noninfectious diseases. However, a molecular mechanism underlying the LPS hyporesponsiveness remains controversial. We here asked whether the TLR4 polymorphism influenced cell surface expression of TLR4/MD-2, ligand-dependent TLR4/MD-2 dimerization or TLR4/ MD-2 responses to a weak agonist monophosphoryl lipid A (MPL). A newly established anti-TLR4 mAb detected D299G/T399I TLR4/MD-2 on Ba/F3 cells whereas a previous anti-TLR4 mAb did not, suggesting that the D299G/T399I polymorphism caused a conformational change in TLR4. Hyporesponsiveness of D299G/T399I TLR4/MD-2 was much more apparent when cells were stimulated with MPL than with lipid A. MPL-dependent TLR4/MD-2 dimerization was impaired by the D299G/T399I polymorphism. The D299G/T399I polymorphism did not alter LPS-binding to soluble TLR4/MD-2, but impaired its dimerization. These results suggest that the D299G/T399I TLR4 polymorphism impairs TLR4/MD-2 responses by altering ligand-dependent dimerization.

Original languageEnglish
Pages (from-to)45-52
Number of pages8
JournalInternational Immunology
Volume25
Issue number1
DOIs
Publication statusPublished - Jan 1 2013
Externally publishedYes

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Toll-Like Receptor 4
Ligands
Dimerization
Lipopolysaccharides
human TLR4 protein
monophosphoryl lipid A
Lipid A

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

Yamakawa, N., Ohto, U., Akashi-Takamura, S., Takahashi, K., Saitoh, S. I., Tanimura, N., ... Miyake, K. (2013). Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A. International Immunology, 25(1), 45-52. https://doi.org/10.1093/intimm/dxs084

Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A. / Yamakawa, Natsuko; Ohto, Umeharu; Akashi-Takamura, Sachiko; Takahashi, Koichiro; Saitoh, Shin Ichiroh; Tanimura, Natsuko; Suganami, Takayoshi; Ogawa, Yoshihiro; Shibata, Takuma; Shimizu, Toshiyuki; Miyake, Kensuke.

In: International Immunology, Vol. 25, No. 1, 01.01.2013, p. 45-52.

Research output: Contribution to journalArticle

Yamakawa, N, Ohto, U, Akashi-Takamura, S, Takahashi, K, Saitoh, SI, Tanimura, N, Suganami, T, Ogawa, Y, Shibata, T, Shimizu, T & Miyake, K 2013, 'Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A', International Immunology, vol. 25, no. 1, pp. 45-52. https://doi.org/10.1093/intimm/dxs084
Yamakawa, Natsuko ; Ohto, Umeharu ; Akashi-Takamura, Sachiko ; Takahashi, Koichiro ; Saitoh, Shin Ichiroh ; Tanimura, Natsuko ; Suganami, Takayoshi ; Ogawa, Yoshihiro ; Shibata, Takuma ; Shimizu, Toshiyuki ; Miyake, Kensuke. / Human TLR4 polymorphism D299G/T399I alters TLR4/MD-2 conformation and response to a weak ligand monophosphoryl lipid A. In: International Immunology. 2013 ; Vol. 25, No. 1. pp. 45-52.
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abstract = "A cell surface heterodimer Toll-like receptor 4 (TLR4)/MD-2 senses lipopolysaccharide (LPS), a principal membrane component of Gram-negative bacteria. LPS binds to MD-2 and induces dimerization of TLR4/MD-2. Dimerized TLR4 activates downstream signaling. TLR4 polymorphism replacing Asp299 with Gly and Thr399 with Ile (D299G/T399I) causes LPS hyporesponsiveness, and is associated with a variety of infectious and noninfectious diseases. However, a molecular mechanism underlying the LPS hyporesponsiveness remains controversial. We here asked whether the TLR4 polymorphism influenced cell surface expression of TLR4/MD-2, ligand-dependent TLR4/MD-2 dimerization or TLR4/ MD-2 responses to a weak agonist monophosphoryl lipid A (MPL). A newly established anti-TLR4 mAb detected D299G/T399I TLR4/MD-2 on Ba/F3 cells whereas a previous anti-TLR4 mAb did not, suggesting that the D299G/T399I polymorphism caused a conformational change in TLR4. Hyporesponsiveness of D299G/T399I TLR4/MD-2 was much more apparent when cells were stimulated with MPL than with lipid A. MPL-dependent TLR4/MD-2 dimerization was impaired by the D299G/T399I polymorphism. The D299G/T399I polymorphism did not alter LPS-binding to soluble TLR4/MD-2, but impaired its dimerization. These results suggest that the D299G/T399I TLR4 polymorphism impairs TLR4/MD-2 responses by altering ligand-dependent dimerization.",
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AU - Saitoh, Shin Ichiroh

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AU - Shimizu, Toshiyuki

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AB - A cell surface heterodimer Toll-like receptor 4 (TLR4)/MD-2 senses lipopolysaccharide (LPS), a principal membrane component of Gram-negative bacteria. LPS binds to MD-2 and induces dimerization of TLR4/MD-2. Dimerized TLR4 activates downstream signaling. TLR4 polymorphism replacing Asp299 with Gly and Thr399 with Ile (D299G/T399I) causes LPS hyporesponsiveness, and is associated with a variety of infectious and noninfectious diseases. However, a molecular mechanism underlying the LPS hyporesponsiveness remains controversial. We here asked whether the TLR4 polymorphism influenced cell surface expression of TLR4/MD-2, ligand-dependent TLR4/MD-2 dimerization or TLR4/ MD-2 responses to a weak agonist monophosphoryl lipid A (MPL). A newly established anti-TLR4 mAb detected D299G/T399I TLR4/MD-2 on Ba/F3 cells whereas a previous anti-TLR4 mAb did not, suggesting that the D299G/T399I polymorphism caused a conformational change in TLR4. Hyporesponsiveness of D299G/T399I TLR4/MD-2 was much more apparent when cells were stimulated with MPL than with lipid A. MPL-dependent TLR4/MD-2 dimerization was impaired by the D299G/T399I polymorphism. The D299G/T399I polymorphism did not alter LPS-binding to soluble TLR4/MD-2, but impaired its dimerization. These results suggest that the D299G/T399I TLR4 polymorphism impairs TLR4/MD-2 responses by altering ligand-dependent dimerization.

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