The Toll-like receptor 4 region Glu24-Pro34 is critical for interaction with MD-2

Biochem Biophys Res Commun. 2005 Mar 11;328(2):586-90. doi: 10.1016/j.bbrc.2005.01.021.

Abstract

Toll-like receptor 4 (TLR4) is a signaling receptor for lipopolysaccharide (LPS) but requires MD-2, a molecule associated with the extracellular TLR4 domain, to respond efficiently to LPS. The purpose of this study was to determine the critical stretch of primary sequence in the TLR4 region involved in MD-2 recognition. TLR4 and TLR4/2a chimera consisting of the TLR4 region Met(1)-Phe(54) and the TLR2 region Ala(53)-Ser(784) were coprecipitated with MD-2, but the deletion mutant TLR4(Delta E24-P34) in which the TLR4 region Glu(24)-Pro(34) was deleted failed to coprecipitate. In agreement with the MD-2 binding, LPS-conjugated beads sedimented TLR4 and TLR4/2a chimera but not TLR2 with MD-2. TLR4(Delta E24-P34) barely coprecipitated with LPS-beads. The cells that had been cotransfected with TLR4(Delta E24-P34) and MD-2 did not induce NF-kappa B activation in response to LPS. These results clearly demonstrate that the amino-terminal TLR4 region of Glu(24)-Pro(34) is critical for MD-2 binding and LPS signaling.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Antigens, Surface / chemistry*
  • Antigens, Surface / metabolism*
  • Binding Sites
  • Carrier Proteins / chemistry*
  • Carrier Proteins / metabolism*
  • Cell Line
  • Humans
  • Kidney / drug effects
  • Kidney / metabolism*
  • Lipopolysaccharides / pharmacology
  • Lymphocyte Antigen 96
  • Membrane Glycoproteins / chemistry*
  • Membrane Glycoproteins / metabolism*
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • NF-kappa B / metabolism*
  • Protein Binding
  • Receptors, Cell Surface / chemistry*
  • Receptors, Cell Surface / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Structure-Activity Relationship
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptors

Substances

  • Antigens, Surface
  • Carrier Proteins
  • LY96 protein, human
  • Lipopolysaccharides
  • Lymphocyte Antigen 96
  • Membrane Glycoproteins
  • NF-kappa B
  • Receptors, Cell Surface
  • Recombinant Proteins
  • TLR2 protein, human
  • TLR4 protein, human
  • Toll-Like Receptor 2
  • Toll-Like Receptor 4
  • Toll-Like Receptors