Purification of granulocyte chemotactic peptide/interleukin-8 reveals N-terminal sequence heterogeneity similar to that of beta-thromboglobulin

Eur J Biochem. 1989 May 1;181(2):337-44. doi: 10.1111/j.1432-1033.1989.tb14729.x.


Stimulated human peripheral blood leukocytes produce a chemotactic factor for granulocytes (granulocyte chemotactic peptide/interleukin-8; GCP/IL-8), which is structurally related to platelet-derived beta-thromboglobulin. Analytically pure CGP/IL-8 and beta-thromboglobulin could be obtained after three purification steps, comprising adsorption to silicic acid, heparin-Sepharose chromatography and ion-exchange chromatography. Although GCP/IL-8 and beta-thromboglobulin had a similar affinity for heparin, they could be separated on a cation-exchange column. Both molecules were heterogeneous in that 6-7-kDa protein doublets were detected upon SDS/PAGE. N-terminal amino acid sequence analysis revealed the presence of six immunologically related but differently truncated polypeptides of beta-thromboglobulin, of which only two corresponded to previously described forms. Similarly, apart from a major polypeptide, five minor species of GCP/IL-8 were detected that also differed by N-terminal truncation. The most processed forms of beta-thromboglobulin and GCP/IL-8 were found to have their N-terminus in that region of the primary structure where a significant similarity between the two molecules starts. GCP/IL-8 was found to be chemotactic for granulocytes with a specific activity of 10(5) units/mg, whereas none of the beta-thromboglobulin species possessed detectable chemotactic activity.

Publication types

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

MeSH terms

  • Amino Acid Sequence
  • Chemotactic Factors / blood
  • Chemotactic Factors / genetics*
  • Chemotactic Factors / isolation & purification
  • Chemotaxis, Leukocyte*
  • Chromatography, Affinity
  • Chromatography, Ion Exchange
  • Electrophoresis, Polyacrylamide Gel
  • Granulocytes / physiology*
  • Humans
  • Interleukin-8
  • Interleukins / blood
  • Interleukins / genetics*
  • Interleukins / isolation & purification
  • Leukocytes / physiology*
  • Molecular Sequence Data
  • Molecular Weight
  • Sequence Homology, Nucleic Acid
  • beta-Thromboglobulin / genetics*
  • beta-Thromboglobulin / isolation & purification


  • Chemotactic Factors
  • Interleukin-8
  • Interleukins
  • beta-Thromboglobulin