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dc.contributor.authorSeo, Een
dc.contributor.authorBlaum, Ben
dc.contributor.authorVargues, Ten
dc.contributor.authorDe Cecco, Men
dc.contributor.authorDeakin, Jon Aen
dc.contributor.authorLyon, Malcolmen
dc.contributor.authorBarran, Pen
dc.contributor.authorCampopiano, Den
dc.contributor.authorUhrín, Den
dc.date.accessioned2011-01-24T11:08:08Z
dc.date.available2011-01-24T11:08:08Z
dc.date.issued2010-12-14
dc.identifier.citationInteraction of human β-defensin 2 (HBD2) with glycosaminoglycans. 2010, 49 (49):10486-95 Biochemistryen
dc.identifier.issn1520-4995
dc.identifier.pmid21062008
dc.identifier.doi10.1021/bi1011749
dc.identifier.urihttp://hdl.handle.net/10541/120252
dc.description.abstractHuman β-defensin 2 (HBD2) is a member of the defensin family of antimicrobial peptides that plays important roles in the innate and adaptive immune system of both vertebrates and invertebrates. In addition to their direct bactericidal action, defensins are also involved in chemotaxis and Toll-like receptor activation. In analogy to chemokine/glycosaminoglycan (GAG) interactions, GAG-defensin complexes are likely to play an important role in chemotaxis and in presenting defensins to their receptors. Using a gel mobility shift assay, we found that HBD2 bound to a range of GAGs including heparin/heparan sulfate (HS), dermatan sulfate (DS), and chondroitin sulfate. We used NMR spectroscopy of (15)N-labeled HBD2 to map the binding sites for two GAG model compounds, a heparin/HS pentasaccharide (fondaparinux sodium; FX) and enzymatically prepared DS hexasaccharide (DSdp6). We identified a number of basic amino acids that form a common ligand binding site, which indicated that these interactions are predominantly electrostatic. The dissociation constant of the [DSdp6-HBD2] complex was determined by NMR spectroscopy to be 5 ± 5 μM. Binding of FX could not be quantified because of slow exchange on the NMR chemical shift time scale. FX was found to induce HBD2 dimerization as evidenced by the analysis of diffusion coefficients, (15)N relaxation, and nESI-MS measurements. The formation of FX-bridged HBD2 dimers exhibited features of a cooperative binding mechanism. In contrast, the complex with DSdp6 was found to be mostly monomeric.
dc.language.isoenen
dc.subject.meshBinding Sites
dc.subject.meshChemotaxis, Leukocyte
dc.subject.meshGlycosaminoglycans
dc.subject.meshHumans
dc.subject.meshMagnetic Resonance Spectroscopy
dc.subject.meshOligosaccharides
dc.subject.meshSpectrometry, Mass, Electrospray Ionization
dc.subject.meshStatic Electricity
dc.subject.meshSulfates
dc.subject.meshbeta-Defensins
dc.titleInteraction of human β-defensin 2 (HBD2) with glycosaminoglycans.en
dc.typeArticleen
dc.contributor.departmentEastChem, School of Chemistry, The University of Edinburgh, King's Buildings, West Mains Road, Edinburgh EH9 3JJ, UK.en
dc.identifier.journalBiochemistryen
html.description.abstractHuman β-defensin 2 (HBD2) is a member of the defensin family of antimicrobial peptides that plays important roles in the innate and adaptive immune system of both vertebrates and invertebrates. In addition to their direct bactericidal action, defensins are also involved in chemotaxis and Toll-like receptor activation. In analogy to chemokine/glycosaminoglycan (GAG) interactions, GAG-defensin complexes are likely to play an important role in chemotaxis and in presenting defensins to their receptors. Using a gel mobility shift assay, we found that HBD2 bound to a range of GAGs including heparin/heparan sulfate (HS), dermatan sulfate (DS), and chondroitin sulfate. We used NMR spectroscopy of (15)N-labeled HBD2 to map the binding sites for two GAG model compounds, a heparin/HS pentasaccharide (fondaparinux sodium; FX) and enzymatically prepared DS hexasaccharide (DSdp6). We identified a number of basic amino acids that form a common ligand binding site, which indicated that these interactions are predominantly electrostatic. The dissociation constant of the [DSdp6-HBD2] complex was determined by NMR spectroscopy to be 5 ± 5 μM. Binding of FX could not be quantified because of slow exchange on the NMR chemical shift time scale. FX was found to induce HBD2 dimerization as evidenced by the analysis of diffusion coefficients, (15)N relaxation, and nESI-MS measurements. The formation of FX-bridged HBD2 dimers exhibited features of a cooperative binding mechanism. In contrast, the complex with DSdp6 was found to be mostly monomeric.


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