Structural studies of heparan sulfate hexasaccharides: new insights into iduronate conformational behavior.
There is a growing opinion that the conformational dynamics within HS chains is critical to their observed biological activities. Investigations into HS conformational dynamics are problematic, given the structural complexity and heterogeneity of HS chains. However, this goal will be more obtainable once we understand the important roles HS sequence/sulfation patterns play in determining the conformational dynamics of iduronate units. This is the first study to compare isomers of N-sulfated oligosaccharides, with respect to the conformational versatility of their internal iduronates. Characterization by NMR spectroscopy of two HS oligosaccharides derived from porcine mucosal HS enabled the measurement of iduronate coupling constants, while under the influence of different flanking saccharide sequences. By fitting our coupling constant data to a new set of theoretical coupling constants, calculated using explicit water molecular dynamic simulations, we are able to offer new insights into the role sequence/sulfation patterns play in influencing iduronate conformational behavior. Fitting of experimental data, using our new theoretically derived coupling constants, suggests that replacement of the N-sulfate group to the reducing side of IdoUA by an N-acetyl group has little effect on the balance of IdoUA conformational equilibrium. Fitting of coupling constants for sequences GlcNS-IdoUA(2S)-GlcNS and GlcNS(6S)-IdoUA(2S)-GlcNS suggests that the flanking 6-O-sulfate group alters the balance of the IdoUA(2S) equilibrium more toward the (2)S0 conformation. There is also the suggestion that a cooperative effect may exist for N- and 6-O sulfation. These observations could be the key to understanding the important regulatory function attributed to 6-O-sulfation within HS chains.
Murphy
Kevin J
KJ
Department of Chemical and Biological Sciences, University of Huddersfield, Huddersfield HD1 3DH, United Kingdom.
McLay
Neil
N
Pye
David A
DA
D000818:Animals; D002236:Carbohydrate Conformation; D006497:Heparan Sulfate / Q000737:chemistry*; D007067:Iduronic Acid / Q000737:chemistry*; D007413:Intestinal Mucosa / Q000737:chemistry; D013058:Mass Spectrometry; D008958:Models, Molecular; D019906:Nuclear Magnetic Resonance, Biomolecular; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification; D013329:Structure-Activity Relationship; D013552:Swine; D013816:Thermodynamics
D006497:Heparan Sulfate; D007067:Iduronic Acid; D009844:Oligosaccharides