Direct determination of the interleukin-6 binding epitope of the interleukin-6 receptor by NMR spectroscopy

J Biol Chem. 2004 Jan 2;279(1):571-6. doi: 10.1074/jbc.M311019200. Epub 2003 Oct 13.

Abstract

All cytokines belonging to the interleukin-6 (IL-6)-type family of cytokines utilize receptors that have a modular build of several immunoglobulin-like and fibronectin type III-like domains. Characteristic of these receptors is a cytokine receptor homology region consisting of two such fibronectin domains defined by a set of four conserved cysteines and a tryptophan-serine-X-tryptophan-serine sequence motif. On target cells, interleukin-6 first binds to its specific receptor and subsequently to a homodimer of the signal transducer protein gp130. The interleukin-6 receptor consists of three extracellular domains. The N-terminal immunoglobulin-like domain is not involved in ligand binding, whereas the third membrane proximal fibronectin-like domain accounts for more than 90% of the binding energy to IL-6. Here, the key residues of this fibronectin-like domain involved in the interaction with IL-6 are described. Chemical shift mapping data with 15N-labeled IL-6R-D3 and unlabeled IL-6 coupled with recent structural data clearly reveal the epitope within the IL-6R-D3 responsible for mediating the high affinity interaction with its cognate cytokine.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Binding Sites
  • Cloning, Molecular
  • Conserved Sequence
  • Cysteine
  • Deuterium
  • Escherichia coli / genetics
  • Interleukin-6 / metabolism*
  • Magnetic Resonance Spectroscopy
  • Models, Molecular
  • Nitrogen Isotopes
  • Protein Conformation
  • Receptors, Interleukin-6 / chemistry*
  • Receptors, Interleukin-6 / metabolism*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism

Substances

  • Interleukin-6
  • Nitrogen Isotopes
  • Receptors, Interleukin-6
  • Recombinant Proteins
  • Deuterium
  • Cysteine