Identification of a glycosaminoglycan binding surface on human interleukin-8

Biochemistry. 1998 Aug 11;37(32):11193-201. doi: 10.1021/bi972867o.


The activation of leukocytes by chemokines is believed to be mediated via binding of chemokines to glycosaminoglycan chains of the extracellular matrix. The binding site on the chemokine interleukin-8 (IL-8) for the glycosaminoglycan heparin has been characterized using a systematic series of site-directed mutants of IL-8 in which the basic residues of the protein have been replaced by alanine. Mutation of K64 and R68 caused the largest decrease in affinity for a heparin Sepharose matrix, with smaller effects seen with mutations of K20, R60, and K67. Heparin-derived disaccharides that could disrupt the IL-8-heparin Sepharose interaction were identified by a competitive binding assay. Heteronuclear NMR spectroscopic titration of 15N-labeled IL-8 with a trisulfated disaccharide revealed a cluster of residues on IL-8 which were perturbed by disaccharide binding. These data identify a heparin-binding surface on IL-8 that includes the C-terminal alpha-helix and the proximal loop around residues 18-23. The heparin-binding site is spatially distinct from the residues involved in receptor binding.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Affinity Labels
  • Binding, Competitive / genetics
  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Disaccharides / metabolism
  • Glycosaminoglycans / chemistry*
  • Glycosaminoglycans / metabolism*
  • Heparin / metabolism
  • Humans
  • Interleukin-8 / chemistry*
  • Interleukin-8 / genetics
  • Interleukin-8 / metabolism*
  • Models, Molecular
  • Mutagenesis, Site-Directed
  • Nuclear Magnetic Resonance, Biomolecular
  • Protein Binding / genetics
  • Sepharose / analogs & derivatives


  • Affinity Labels
  • Disaccharides
  • Glycosaminoglycans
  • Interleukin-8
  • heparin-sepharose
  • Heparin
  • Sepharose