A 3D-structural model of unsulfated chondroitin from high-field NMR: 4-sulfation has little effect on backbone conformation |
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Authors: | Benedict M Sattelle Ian S Roberts |
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Institution: | a Manchester Interdisciplinary Biocentre, 131 Princess Street, Manchester, M1 7DN, UK b Michael Smith Building, Oxford Road, Manchester, M13 9PT, UK |
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Abstract: | The glycosaminoglycan chondroitin sulfate is essential in human health and disease but exactly how sulfation dictates its 3D-strucutre at the atomic level is unclear. To address this, we have purified homogenous oligosaccharides of unsulfated chondroitin (with and without 15N-enrichment) and analysed them by high-field NMR to make a comparison published chondroitin sulfate and hyaluronan 3D-structures. The result is the first full assignment of the tetrasaccharide and an experimental 3D-model of the hexasaccharide (PDB code 2KQO). In common with hyaluronan, we confirm that the amide proton is not involved in strong, persistent inter-residue hydrogen bonds. However, in contrast to hyaluronan, a hydrogen bond is not inferred between the hexosamine OH-4 and the glucuronic acid O5 atoms across the β(1→3) glycosidic linkage. The unsulfated chondroitin bond geometry differs slightly from hyaluronan by rotation about the β(1→3) ψ dihedral (as previously predicted by simulation), while the β(1→4) linkage is unaffected. Furthermore, comparison shows that this glycosidic linkage geometry is similar in chondroitin-4-sulfate. We therefore hypothesise that both hexosamine OH-4 and OH-6 atoms are solvent exposed in chondroitin, explaining why it is amenable to sulfation and hyaluronan is not, and also that 4-sulfation has little effect on backbone conformation. Our conclusions exemplify the value of the 3D-model presented here and progress our understanding of glycosaminoglycan molecular properties. |
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Keywords: | CN chondroitin (unsulfated) COSY correlation spectroscopy CS chondriotin sulfate CSPG CS proteoglycan GAG glycosaminoglycan GlcA d-glucuronic acid" target="_blank">d-glucuronic acid GalNAc d-galactosamine" target="_blank">N-acetyl-d-galactosamine GlcNAc d-glucosamine" target="_blank">N-acetyl-d-glucosamine HA hyaluronan HMBC heteronuclear multiple bond coherence HSQC heteronuclear single quantum coherence NMR nuclear magnetic resonance NOESY nuclear Overhauser effect spectroscopy PDB Protein data bank PG proteoglycan |
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