Different affinities of glycosaminoglycan oligosaccharides for monomeric and dimeric interleukin-8: a model for chemokine regulation at inflammatory sites.

2.50
Hdl Handle:
http://hdl.handle.net/10541/83580
Title:
Different affinities of glycosaminoglycan oligosaccharides for monomeric and dimeric interleukin-8: a model for chemokine regulation at inflammatory sites.
Authors:
Goger, Birgit; Halden, Yvonne; Rek, Angelika; Mösl, Roland; Pye, David A; Gallagher, John T; Kungl, Andreas J
Abstract:
The binding of interleukin-8 (IL-8) to heparan sulfate (HS) proteoglycans on the surface of endothelial cells is crucial for the recruitment of neutrophils to an inflammatory site. Fluorescence anisotropy measurements yielded an IL-8 dimerization constant of 120 nM. The binding affinities, obtained by isothermal fluorescence titration, of size-defined heparin and HS oligosaccharides to the chemokine were found to depend on the oligomerization state of IL-8: high affinity was detected for monomeric and low affinity was detected for dimeric IL-8, referring to a self-regulatory mechanism for its chemoattractant effect. The highest affinity for monomeric IL-8 was detected for the HS octamer with a K(d) < 5 nM whereas the dissociation constants of dimeric IL-8 were found in the medium micromolar range. No indication for increasing affinities for monomeric IL-8 with increasing oligosaccharide chain length was found. Instead, a periodic pattern was obtained for the dissociation constants of the GAG oligosaccharides with respect to chain length, referring to optimum and least optimum chain lengths for IL-8 binding. GAG disaccharides were identified to be the minimum length for chemokine binding. Conformational changes of the dimeric chemokine, determined using CD spectroscopy, were detected only for the IL-8/HS complexes and not for heparin, pointing to an HS-induced activation of the chemokine with respect to receptor binding. Thermal unfolding of IL-8 yielded a single transition at 56 degrees C which was completely prevented by the presence of undigested HS or heparin, indicating structural stabilization, thereby prolonging the biological effect of the chemokine.
Affiliation:
Institute of Pharmaceutical Chemistry, Protein Chemistry and Biophysics Group, University of Graz, Universitätsplatz 1, A-8010 Graz, Austria.
Citation:
Different affinities of glycosaminoglycan oligosaccharides for monomeric and dimeric interleukin-8: a model for chemokine regulation at inflammatory sites. 2002, 41 (5):1640-6 Biochemistry
Journal:
Biochemistry
Issue Date:
5-Feb-2002
URI:
http://hdl.handle.net/10541/83580
DOI:
10.1021/bi011944j
PubMed ID:
11814358
Type:
Article
Language:
en
ISSN:
0006-2960
Appears in Collections:
All Paterson Institute for Cancer Research

Full metadata record

DC FieldValue Language
dc.contributor.authorGoger, Birgiten
dc.contributor.authorHalden, Yvonneen
dc.contributor.authorRek, Angelikaen
dc.contributor.authorMösl, Rolanden
dc.contributor.authorPye, David Aen
dc.contributor.authorGallagher, John Ten
dc.contributor.authorKungl, Andreas Jen
dc.date.accessioned2009-10-05T15:48:35Z-
dc.date.available2009-10-05T15:48:35Z-
dc.date.issued2002-02-05-
dc.identifier.citationDifferent affinities of glycosaminoglycan oligosaccharides for monomeric and dimeric interleukin-8: a model for chemokine regulation at inflammatory sites. 2002, 41 (5):1640-6 Biochemistryen
dc.identifier.issn0006-2960-
dc.identifier.pmid11814358-
dc.identifier.doi10.1021/bi011944j-
dc.identifier.urihttp://hdl.handle.net/10541/83580-
dc.description.abstractThe binding of interleukin-8 (IL-8) to heparan sulfate (HS) proteoglycans on the surface of endothelial cells is crucial for the recruitment of neutrophils to an inflammatory site. Fluorescence anisotropy measurements yielded an IL-8 dimerization constant of 120 nM. The binding affinities, obtained by isothermal fluorescence titration, of size-defined heparin and HS oligosaccharides to the chemokine were found to depend on the oligomerization state of IL-8: high affinity was detected for monomeric and low affinity was detected for dimeric IL-8, referring to a self-regulatory mechanism for its chemoattractant effect. The highest affinity for monomeric IL-8 was detected for the HS octamer with a K(d) < 5 nM whereas the dissociation constants of dimeric IL-8 were found in the medium micromolar range. No indication for increasing affinities for monomeric IL-8 with increasing oligosaccharide chain length was found. Instead, a periodic pattern was obtained for the dissociation constants of the GAG oligosaccharides with respect to chain length, referring to optimum and least optimum chain lengths for IL-8 binding. GAG disaccharides were identified to be the minimum length for chemokine binding. Conformational changes of the dimeric chemokine, determined using CD spectroscopy, were detected only for the IL-8/HS complexes and not for heparin, pointing to an HS-induced activation of the chemokine with respect to receptor binding. Thermal unfolding of IL-8 yielded a single transition at 56 degrees C which was completely prevented by the presence of undigested HS or heparin, indicating structural stabilization, thereby prolonging the biological effect of the chemokine.en
dc.language.isoenen
dc.subject.meshAnimals-
dc.subject.meshBinding Sites-
dc.subject.meshCell Line-
dc.subject.meshCircular Dichroism-
dc.subject.meshDimerization-
dc.subject.meshGlycosaminoglycans-
dc.subject.meshHeparin-
dc.subject.meshHeparitin Sulfate-
dc.subject.meshHot Temperature-
dc.subject.meshHumans-
dc.subject.meshInflammation-
dc.subject.meshInterleukin-8-
dc.subject.meshKinetics-
dc.subject.meshModels, Chemical-
dc.subject.meshOligosaccharides-
dc.subject.meshProtein Binding-
dc.subject.meshProtein Denaturation-
dc.subject.meshProtein Folding-
dc.subject.meshProtein Structure, Quaternary-
dc.subject.meshSpectrometry, Fluorescence-
dc.subject.meshSwine-
dc.titleDifferent affinities of glycosaminoglycan oligosaccharides for monomeric and dimeric interleukin-8: a model for chemokine regulation at inflammatory sites.en
dc.typeArticleen
dc.contributor.departmentInstitute of Pharmaceutical Chemistry, Protein Chemistry and Biophysics Group, University of Graz, Universitätsplatz 1, A-8010 Graz, Austria.en
dc.identifier.journalBiochemistryen

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