Regulatory roles for MD-2 and TLR4 in ligand-induced receptor clustering

Makiko Kobayashi, Shin Ichiroh Saitoh, Natsuko Tanimura, Koichiro Takahashi, Kiyoshi Kawasaki, Masahiro Nishijima, Yukari Fujimoto, Koichi Fukase, Sachiko Akashi-Takamura, Kensuke Miyake

Research output: Contribution to journalArticle

127 Citations (Scopus)

Abstract

LPS, a principal membrane component in Gram-negative bacteria, is recognized by a receptor complex consisting of TLR4 and MD-2. MD-2 is an extracellular molecule that is associated with the extracellular domain of TLR4 and has a critical role in LPS recognition. MD-2 directly interacts with LPS, and the region from Phe119 to Lys132 (Arg132 in mice) has been shown to be important for interaction between LPS and TLR4/MD-2. With mouse MD-2 mutants, we show in this study that Gly59 was found to be a novel critical amino acid for LPS binding outside the region 119-132. LPS signaling is thought to be triggered by ligand-induced TLR4 clustering, which is also regulated by MD-2. Little is known, however, about a region or an amino acid in the MD-2 molecule that regulates ligand-induced receptor clustering. MD-2 mutants substituting alanine for Phe126 or Gly 129 impaired LPS-induced TLR4 clustering, but not LPS binding to TLR4/MD-2, demonstrating that ligand-induced receptor clustering is differentially regulated by MD-2 from ligand binding. We further show that dissociation of ligand-induced receptor clustering and of ligand-receptor interaction occurs in a manner dependent on TLR4 signaling and requires endosomal acidification. These results support a principal role for MD-2 in LPS recognition.

Original languageEnglish
Pages (from-to)6211-6218
Number of pages8
JournalJournal of Immunology
Volume176
Issue number10
Publication statusPublished - 2006 May 15
Externally publishedYes

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Cluster Analysis
Ligands
Amino Acids
Gram-Negative Bacteria
Alanine
Membranes

ASJC Scopus subject areas

  • Immunology

Cite this

Kobayashi, M., Saitoh, S. I., Tanimura, N., Takahashi, K., Kawasaki, K., Nishijima, M., ... Miyake, K. (2006). Regulatory roles for MD-2 and TLR4 in ligand-induced receptor clustering. Journal of Immunology, 176(10), 6211-6218.

Regulatory roles for MD-2 and TLR4 in ligand-induced receptor clustering. / Kobayashi, Makiko; Saitoh, Shin Ichiroh; Tanimura, Natsuko; Takahashi, Koichiro; Kawasaki, Kiyoshi; Nishijima, Masahiro; Fujimoto, Yukari; Fukase, Koichi; Akashi-Takamura, Sachiko; Miyake, Kensuke.

In: Journal of Immunology, Vol. 176, No. 10, 15.05.2006, p. 6211-6218.

Research output: Contribution to journalArticle

Kobayashi, M, Saitoh, SI, Tanimura, N, Takahashi, K, Kawasaki, K, Nishijima, M, Fujimoto, Y, Fukase, K, Akashi-Takamura, S & Miyake, K 2006, 'Regulatory roles for MD-2 and TLR4 in ligand-induced receptor clustering', Journal of Immunology, vol. 176, no. 10, pp. 6211-6218.
Kobayashi M, Saitoh SI, Tanimura N, Takahashi K, Kawasaki K, Nishijima M et al. Regulatory roles for MD-2 and TLR4 in ligand-induced receptor clustering. Journal of Immunology. 2006 May 15;176(10):6211-6218.
Kobayashi, Makiko ; Saitoh, Shin Ichiroh ; Tanimura, Natsuko ; Takahashi, Koichiro ; Kawasaki, Kiyoshi ; Nishijima, Masahiro ; Fujimoto, Yukari ; Fukase, Koichi ; Akashi-Takamura, Sachiko ; Miyake, Kensuke. / Regulatory roles for MD-2 and TLR4 in ligand-induced receptor clustering. In: Journal of Immunology. 2006 ; Vol. 176, No. 10. pp. 6211-6218.
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