Characterization of a domain of a human type I interferon receptor protein involved in ligand binding

J Interferon Cytokine Res. 1995 Mar;15(3):205-11. doi: 10.1089/jir.1995.15.205.


Two monoclonal antibodies that recognize different epitopes of the extracellular domain of one of the proteins that constitute the type I interferon receptor were used to delineate the interferon binding site. Antibody 64G12 both inhibits the binding of radiolabeled interferon-alpha 2 and IFN-alpha 8 to their cell surface receptors and neutralizes the antiviral and antiproliferative actions of all the type I interferons tested, including IFN-beta, IFN-omega, and human leukocyte IFN, a mixture of different interferon-alpha isotypes. Antibody 34F10 recognizes the type I interferon receptor with an affinity similar to that of the MAb 64G12 but does not inhibit either the binding or the biologic activity of any of the type I interferons tested. Both antibodies recognize a protein of 105 +/- 5 kD from either Daudi or Ly28 cells. Immunoprecipitation following surface iodination demonstrated that the neutralizing MAb recognizes a protein of 105 kD and the nonneutralizing MAb a protein of 110 kD in extracts of Daudi cells. A second less intense band was also detected by both antibodies. Cross-linking of IFN-alpha 2 to its receptor before immunoprecipitation prevented the neutralizing antibody from immunoprecipitating the receptor protein, but the nonneutralizing MAb was still able to recognize a 140 kD protein corresponding to the cross-linked interferon-receptor protein complex. Thus, an interferon binding domain appears to be localized in a region between amino acids 23 and 229 of the extracellular domain of a transmembrane protein that forms part of the type I interferon receptor complex containing the epitopes recognized by each antibody.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Binding Sites
  • Blotting, Western
  • Cross-Linking Reagents
  • Epitopes / chemistry
  • Epitopes / immunology
  • Humans
  • Interferon Type I / chemistry*
  • Interferon Type I / immunology
  • Ligands
  • Membrane Proteins
  • Neutralization Tests
  • Precipitin Tests
  • Receptor, Interferon alpha-beta
  • Receptors, Interferon / chemistry*
  • Receptors, Interferon / immunology
  • Tumor Cells, Cultured


  • Antibodies, Monoclonal
  • Cross-Linking Reagents
  • Epitopes
  • Interferon Type I
  • Ligands
  • Membrane Proteins
  • Receptors, Interferon
  • Receptor, Interferon alpha-beta