Rational design of a receptor super-antagonist of human interleukin-6

EMBO J. 1994 Dec 15;13(24):5863-70. doi: 10.1002/j.1460-2075.1994.tb06931.x.

Abstract

Interleukin-6 (IL-6) is a differentiation and growth factor for a variety of cell types and its excessive production plays a major role in the pathogenesis of multiple myeloma and post-menopausal osteoporosis. IL-6, a four-helix bundle cytokine, is believed to interact sequentially with two transmembrane receptors, the low-affinity IL-6 receptor (IL-6R alpha) and the signal transducer gp130, via distinct binding sites. In this paper we show that combined mutations in the predicted A and C helices, previously suggested to establish contacts with gp130, give rise to variants with no bioactivity but unimpaired binding to IL-6R alpha. These mutants behave as full and selective IL-6 receptor antagonists on a variety of human cell lines. Furthermore, a bifacial mutant was generated (called IL-6 super-antagonist) in which the antagonist mutations were combined with amino acid substitutions in the predicted D helix that increase binding for IL-6R alpha. The IL-6 super-antagonist has no bioactivity, but improved first receptor occupancy and, therefore, fully inhibits the wild-type cytokine at low dosage. The demonstration of functionally independent receptor binding sites on IL-6 suggests that it could be possible to design super-antagonists of other helical cytokines which drive the assembly of structurally related multisubunit receptor complexes.

Publication types

  • Comparative Study

MeSH terms

  • Carcinoma, Hepatocellular / metabolism
  • Dose-Response Relationship, Drug
  • Drug Design
  • Humans
  • Interleukin-6 / analogs & derivatives*
  • Interleukin-6 / genetics
  • Interleukin-6 / pharmacology
  • Liver Neoplasms / metabolism
  • Models, Molecular
  • Multiple Myeloma / metabolism
  • Mutagenesis, Site-Directed
  • Protein Structure, Secondary
  • Receptors, Interleukin / antagonists & inhibitors*
  • Receptors, Interleukin-6
  • Sensitivity and Specificity
  • Tumor Cells, Cultured

Substances

  • Interleukin-6
  • Receptors, Interleukin
  • Receptors, Interleukin-6