Complete assignment of hyaluronan oligosaccharides up to hexasaccharides

Blundell, Charles; Reed, Michelle A.C.; Almond, Andrew

doi:10.1016/j.carres.2006.09.023

Cited by 39 publications

(32 citation statements)

References 32 publications

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“…The abbreviation K5 n denotes a K5 oligosaccharide of n monosaccharide residues with ⌬ 4,5-GlcA at the non-reducing terminus. [Blundell et al, 2006b]. [ 1 H, 13 C]-HSQC spectra with and without 13 C-broadband decoupling disabled during acquisition were recorded at 13 C-natural abundance [Blundell et al, 2006b].…”

Section: Methodsmentioning

confidence: 99%

“…[Blundell et al, 2006b]. [ 1 H, 13 C]-HSQC spectra with and without 13 C-broadband decoupling disabled during acquisition were recorded at 13 C-natural abundance [Blundell et al, 2006b]. Amide temperature coefficients of chemical shifts ( ⌬ ␦ / ⌬ T) were derived from [ 1 H]-1D and [ 1 H, 15 N]-HSQC datasets recorded at 15 N-natural abundance and 750 MHz, as described previously [Blundell and Almond, 2007].…”

Section: Methodsmentioning

confidence: 99%

“…3 ) and, as in D-GlcNAc [Blundell et al, 2006b] changes in the rotamer populations (i.e., all ϳ 1: 1 gg : gt ) [Haasnoot et al, 1980]. Interestingly, H-6 proR and H-6 proS always have isochronous chemical shifts in the GlcNAc ring adjacent to a non-reducing terminal ⌬ 4,5GlcA ring (see table 1 and fig.…”

Section: Conformation-dependent Coupling Constants In K5 Oligosaccharmentioning

confidence: 99%

“…4 C 1 chairs) [Blundell et al, 2006b]. Moreover, these large values show that the K5 oligosaccharides have the correct ring stereochemistry (i.e., a glucose base [Vann et al, 1981]), which is important because other stereoisomers (e.g., idose base, galactose base) may not have been detected or resolved by the anion-exchange FPLC or MALDI-ToF analyses.…”

Section: Conformation-dependent Coupling Constants In K5 Oligosaccharmentioning

confidence: 99%

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Investigating the Molecular Basis for the Virulence of <i>Escherichia coli</i> K5 by Nuclear Magnetic Resonance Analysis of the Capsule Polysaccharide

Blundell

Roberts²,

Sheehan³

et al. 2009

Microb Physiol

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The capsular polysaccharide of Escherichia coli K5 has been hypothesised to promote virulence through its molecular mimicry of host heparan sulphate. To test this hypothesis, we have produced pure oligosaccharides from K5 capsular polysaccharide and investigated their conformational properties with ultra-high-field nuclear magnetic resonance (NMR) (900 MHz). Ultra-high-field affords a significant resolution enhancement over previous studies and allowed a full-atomic assignment of the K5 hexasaccharide for the first time. All carbohydrate rings adopt a ⁴C₁ conformation, the amide sidechains have a trans orientation and the hydroxymethyl group is freely exposed to bulk solvent. Initial models of the glycosidic linkage conformation based upon simple interpretation of NOE cross-peaks suggests that the β1→4 linkage adopts a 3D geometry of φ ≈ 60°, ψ ≈ 0° and the α1→4 linkage prefers φ ≈ –30°, ψ ≈ –30° (φ and ψ being defined by dihedral angles involving linkage protons). In this conformation the overall molecular geometries of K5 polysaccharide, heparan sulphate and even fully-sulphated heparin are remarkably similar. These results substantiate the hypothesis that the K5 capsular polysaccharide confers virulence to E. coli K5 by being a 3D molecular mimetic of host heparan sulphate, helping it to evade detection by the mammalian immune system.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Conformation-dependent Coupling Constants In K5 Oligosaccharmentioning

confidence: 99%

Section: Conformation-dependent Coupling Constants In K5 Oligosaccharmentioning

confidence: 99%

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Investigating the Molecular Basis for the Virulence of <i>Escherichia coli</i> K5 by Nuclear Magnetic Resonance Analysis of the Capsule Polysaccharide

Blundell

Roberts²,

Sheehan³

et al. 2009

Microb Physiol

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“…Unfortunately, this method results in nearly identical retention times for the HA and NAH oligosaccharides, and since they have identical masses, it is not possible to differentiate these oligosaccharides. Most laboratories routinely rely on nuclear magnetic resonance (NMR) spectroscopy, requiring relatively large amounts of samples, to distinguish HA and NAH oligosaccharides [26,27]. These two series of oligosaccharides can also be differentiated based on their susceptibilities to specific polysaccharide degrading enzymes, where the degradation can be detected by ultraviolet spectroscopy (UV), HPLC, thin-layer chromatography (TLC), polyacrylamide gel electrophoresis, or change in viscosity.…”

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confidence: 99%

Tandem MS can distinguish hyaluronic acid from N-acetylheparosan

Zhang

Xie

Liu³

et al. 2008

J. Am. Soc. Mass Spectrom.

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Isobaric oligosaccharides enzymatically prepared from hyaluronic acid (HA) and N-acetylheparosan (NAH), were distinguished using tandem mass spectrometry. The only difference between the two series of oligosaccharides was the linkage pattern (in HA 1¡3 and in NAH 1¡4) between glucuronic acid and N-acetylglucosamine residues. Tandem mass spectrometry afforded spectra in which glycosidic cleavage fragment ions were observed for both HA and NAH oligosaccharides. Cross-ring cleavage ions 0,2 A n and 0,2 A n -h (n is even number) were observed only in GlcNAc residues of NAH oligosaccharides. One exception was an 0,2 A 2 ion fragment observed for the disaccharide from HA. These cross-ring cleavage fragment ions are useful to definitively distinguish HA and NAH oligosaccharides. -3]. GAGs are divided into four main categories: hyaluronic acid or hyaluronan (HA), chondroitin/dermatan sulfate, Nacetylheparosan (NAH)/heparan sulfate/heparin, and keratan sulfate, based on their monosaccharide composition and the configuration and position of the glycosidic bonds between these monosaccharides. The specificity of the interactions between GAGs and proteins results from structural diversity of GAGs defined by their size, saccharide composition and sequence, charge density [4,5]. Thus, understanding the structure of a GAG is essential in understanding its activity and biological functions. HA and NAH are biosynthesized by both prokaryotic and eukaryotic cells. HA is copolymer linked polymer of ␤-1,4 D-glucuronic acid (GlcA) and ␤-1,3 D-N-acetylglucosamine (GlcNAc) [6]. NAH is a copolymer of ␤-1,4 GlcA and ␣-1,4 GlcNAc and the biosynthetic precursor of the mammalian GAGs, heparin and heparan sulfate [7]. HA can be prepared by bacterial fermentation of Streptococcus zooepidemicus [8], or enzymatically using biosynthetic enzymes prepared from Pasturella multocida [9]. Bacteria use HA as camouflage to enhance their ability to infect higher animals [10]. Similarly, Escherichia coli strain K5 produces capsular polysaccharide comprised of NAH [11]. NAH is also important as it represents a considerable portion of the sequence of mammalian heparan sulfate [12,13]. HA and NAH are the two simplest GAGs since neither has sulfo groups and, thus, consist of only a single sequence (Figure 1). HA and NAH have very similar structures, differing only in the position of their glycosidic linkage between GlcA and GlcNAc. This difference in structure results from very different biosynthesis pathways for HA and NAH [14 -17] and gives each polysaccharide unique biological functions and distinctive structure-activity relationships [18].The oligosaccharides prepared from GAGs by controlled enzymatic depolymerization are often studied to define the minimum structural requirements for biological activity [19,20]. ESI-MS is particularly useful to monitor these GAG-derived oligosaccharides due to its soft ionization [21,22]. LC-ESI-MS, relying on reversedphase ion-pairing (RPIP)-high-performance liquid chromatography (HPLC), has employed volatile ion-pai...

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Chemoenzymatic Synthesis of Nonasulfated Tetrahyaluronan with a Paramagnetic Tag for Studying Its Complex with Interleukin‐10

Köhling

Künze

Lemmnitzer

et al. 2016

Chemistry A European J

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Implants and artificial biomaterials containing sulfated hyaluronans have been shown to improve the healing of injured skin and bones. It is hypothesized that these effects are mediated by the binding of sulfated glycosaminoglycans (GAGs) to growth factors and cytokines, resulting in the sequestering of proteins to the wound healing site and in modulated protein activity. Given that no direct synthetic access to sulfated oligohyaluronans has been available, little is known about their protein binding and the structure of the resulting protein complexes. Here, the chemoenzymatic preparation of oligohyaluronans on the gram scale is described. Oligohyaluronans are converted into anomeric azides at the reducing end, enabling the attachment of analytical labels through an anomeric ligation reaction. A nonasulfated tetrahyaluronan-ethylenediaminetetraacetic acid derivative has been produced and used as a paramagnetic tag for the elucidation of the complex of this ligand with interleukin-10 using paramagnetic relaxation enhancement NMR analysis. The metal ion position is resolved with 1.0 Å, enabling a refined structural model of the complex.

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Complete assignment of hyaluronan oligosaccharides up to hexasaccharides

Cited by 39 publications

References 32 publications

Investigating the Molecular Basis for the Virulence of <i>Escherichia coli</i> K5 by Nuclear Magnetic Resonance Analysis of the Capsule Polysaccharide

Investigating the Molecular Basis for the Virulence of <i>Escherichia coli</i> K5 by Nuclear Magnetic Resonance Analysis of the Capsule Polysaccharide

Tandem MS can distinguish hyaluronic acid from N-acetylheparosan

Chemoenzymatic Synthesis of Nonasulfated Tetrahyaluronan with a Paramagnetic Tag for Studying Its Complex with Interleukin‐10

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