Interleukin-8 binds to syndecan-2 on human endothelial cells
| Autor: | Werner Atzenhofer, Astrid Wabnig, Angelika Rek, Laszlo Szilak, Andreas J. Kungl, Yvonne Halden |
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| Rok vydání: | 2004 |
| Předmět: |
Glycosylation
Immunoprecipitation behavioral disciplines and activities Biochemistry Protein Structure Secondary Umbilical vein Syndecan 1 Humans Interleukin 8 Molecular Biology Cells Cultured Glycosaminoglycan binding Binding Sites Membrane Glycoproteins biology Tumor Necrosis Factor-alpha Chemistry Interleukin-8 Cell Biology Transfection Molecular biology carbohydrates (lipids) Proteoglycan Ectodomain embryonic structures biology.protein Proteoglycans Endothelium Vascular Syndecan-2 Research Article |
| Zdroj: | Biochemical Journal. 377:533-538 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj20030729 |
| Popis: | Application of reverse transcription-PCR to total RNA prepared from TNF-alpha (tumour necrosis factor-alpha)-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-alpha-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 [Syn2(ect)(+HS)] and non-glycosylated [Syn2(ect)(-HS)] forms of Syn2(ect) (the syndecan-2 ectodomain) were purified from a stably transfected human cell line and from a bacterial expression system respectively. By CD spectroscopy, Syn2(ect) was found to adopt an all-beta secondary structure. The dissociation constant of Syn2(ect)(+HS) with respect to interleukin-8 binding was determined by isothermal fluorescence titrations to be 23 nM. Despite its lack of HS chains, Syn2(ect)(-HS) exhibited significant binding to the chemokine, with a K (d) of >1 microM. Thus, in addition to glycosaminoglycan binding, protein-protein contacts might also contribute to the chemokine-proteoglycan interaction. |
| Databáze: | OpenAIRE |
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