Identification of the domain in the human interleukin-11 receptor that mediates ligand binding

J Mol Biol. 2001 Feb 16;306(2):263-74. doi: 10.1006/jmbi.2000.4387.

Abstract

The interleukin-11 receptor (IL-11R) belongs to the hematopoietic receptor superfamily. The functional receptor complex comprises IL-11, IL-11R and the signal-transducing subunit gp130. The extracellular part of the IL-11R consists of three domains: an N-terminal immunoglobulin-like domain, D1, and two fibronectin-type III-like (FNIII) domains and D2 and D3. The two FNIII domains comprise the cytokine receptor-homology region defined by a set of four conserved cysteine residues in the N-terminal domain (D2) and a WSXWS sequence motif in the C-terminal domain (D3). We investigated the structural and functional role of the third extracellular receptor domain of IL-11R. A molecular model of the human IL-11/IL-11R complex allowed the identification of amino acid residues in IL-11R to be involved in ligand binding. Most of them were located in the third extracellular domain, which therefore should be able to bind with high affinity to IL-11. To prove this prediction, domain D3 of the IL-11R was expressed in Escherichia coli, refolded and purified. For structural characterization, circular dichroism, fluorescence and NMR spectroscopy were used. By plasmon resonance experiments, we show that the ligand-binding capacity of this domain is as high as that one for the whole receptor. These results provide a basis for further structural investigations that could be used for the rational design of potential agonists and antagonists essential in human therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Circular Dichroism
  • Conserved Sequence
  • Cysteine / metabolism
  • Fluorescence Polarization
  • Humans
  • Interleukin-11 Receptor alpha Subunit
  • Kinetics
  • Ligands
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Denaturation
  • Protein Folding
  • Protein Renaturation
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Receptors, Interleukin / chemistry*
  • Receptors, Interleukin / genetics
  • Receptors, Interleukin / isolation & purification
  • Receptors, Interleukin / metabolism*
  • Receptors, Interleukin-11
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Sequence Alignment
  • Surface Plasmon Resonance
  • Temperature
  • Thermodynamics

Substances

  • IL11RA protein, human
  • Interleukin-11 Receptor alpha Subunit
  • Ligands
  • Receptors, Interleukin
  • Receptors, Interleukin-11
  • Recombinant Proteins
  • Cysteine