Interleukin-8 binds to syndecan-2 on human endothelial cells

Yvonne Halden, Angelika Rek, Werner Atzenhofer, Laszlo Szilak, Astrid Wabnig, Andreas J. Kungl

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Application of reverse transcription-PCR to total RNA prepared from TNF-α (tumour necrosis factor-α)-stimulated HUVECs (human umbilical vein endothelial cells) revealed that the syndecan-2 mRNA was up-regulated by this inflammatory stimulus. By immunoprecipitation using an anti-syndecan-2 antibody on TNF-α-stimulated HUVEC lysates, inflammation-induced interleukin-8 was found to be an interaction partner of this HS (heparan sulphate) proteoglycan, but not of any other syndecan on these cells. The glycosylated [Syn2ect(+HS)] and non-glycosylated [Syn2 ect(-HS)] forms of Syn2ect (the syndecan-2 ectodomain) were purified from a stably transfected human cell line and from a bacterial expression system respectively. By CD spectroscopy, Syn2ect was found to adopt an all-β secondary structure. The dissociation constant of Syn2ect(+ HS) with respect to interleukin-8 binding was determined by isothermal fluorescence titrations to be 23 nM. Despite its lack of HS chains, Syn2ect(- HS) exhibited significant binding to the chemokine, with a Kd of > 1 μM. Thus, in addition to glycosaminoglycan binding, protein-protein contacts might also contribute to the chemokine-proteoglycan interaction.

Original languageEnglish
Pages (from-to)533-538
Number of pages6
JournalBiochemical Journal
Volume377
Issue number2
DOIs
Publication statusPublished - Jan 15 2004

Keywords

  • Chemokine
  • Heparan sulphate
  • Interleukin-8
  • Proteoglycan
  • Syndecan-2

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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  • Cite this

    Halden, Y., Rek, A., Atzenhofer, W., Szilak, L., Wabnig, A., & Kungl, A. J. (2004). Interleukin-8 binds to syndecan-2 on human endothelial cells. Biochemical Journal, 377(2), 533-538. https://doi.org/10.1042/BJ20030729