Saturation mutagenesis of the human interleukin 6 receptor-binding site: implications for its three-dimensional structure

Proc Natl Acad Sci U S A. 1993 May 1;90(9):4067-71. doi: 10.1073/pnas.90.9.4067.

Abstract

Interleukin 6 is a 184-aa polypeptide postulated to belong to the class of helical cytokines. We built a three-dimensional model of human interleukin 6 based on the similarity of its hydrophobicity pattern with that of other cytokines and on the x-ray structure of growth hormone, interleukin 2, interleukin 4, interferon beta, and granulocyte-macrophage colony-stimulating factor. The resulting model is a bundle of four alpha-helices and suggests possible alternative conformations for the 9 C-terminal amino acids; in this region, the importance of Arg-182 and Met-184 for biological activity has been demonstrated [Lutticken, C., Kruttgen, A., Moller, C., Heinrich, P.C. & Rose-John, S. (1991) FEBS Lett. 282, 265-267]. Therefore, we generated a large collection of single-amino acid variants in residues 175-181. Analysis of their biological activity in two systems and the receptor binding properties of a subset of the mutants indicates that the entire region is involved in forming the receptor binding surface and supports the hypothesis that this region does not assume an alpha-helical conformation. Remarkably, we also found a mutant with receptor affinity and biological activity much higher than wild type; the potential therapeutical value of this finding is discussed.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Base Sequence
  • Binding Sites
  • Biological Assay
  • Cytokines / chemistry
  • Cytokines / genetics
  • Humans
  • Interleukin-6 / chemistry*
  • Interleukin-6 / genetics
  • Interleukin-6 / metabolism*
  • Kinetics
  • Models, Molecular
  • Molecular Sequence Data
  • Mutagenesis, Site-Directed
  • Protein Structure, Secondary*
  • Receptors, Immunologic / metabolism*
  • Receptors, Interleukin-6

Substances

  • Cytokines
  • Interleukin-6
  • Receptors, Immunologic
  • Receptors, Interleukin-6