1H Nuclear Magnetic Resonance Spectroscopic Analysis for Determination of Glucuronic and Iduronic Acids in Dermatan Sulfate, Heparin, and Heparan Sulfate

Sudo, Masahiro; Satô, Kenji; Chaidedgumjorn, Amornrut; Toyoda, Hidenao; Toida, Toshihiko; Imanari, Toshio

doi:10.1006/abio.2001.5296

Cited by 36 publications

(17 citation statements)

References 40 publications

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“…In particular, signals labeled c, e and i at 3.3 ppm, 3.6-3.7 ppm and 4.4 ppm, correspond to individual protons in glucuronic acid, while the signal at 5.1 ppm labeled j corresponds to iduronic acid. The spectra of HS clearly demonstrated very low level of iduronic acid content in all HS samples examined [25]. This is an important observation since disaccharide analysis using heparin lyases can not be used to definitively distinguish between glucuronic and iduronic acid.…”

Section: Structural Characterization Of Organs Hs Using 1 H-nmr Spectmentioning

confidence: 91%

Isolation and characterization of heparan sulfate from various murine tissues

et al. 2006

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Heparan sulfate (HS), is a proteoglycan (PG) found both in the extracellular matrix and on cell surface. It may represent one of the most biologically important glycoconjugates, playing an essential role in a variety of different events at molecular level. The publication of the mouse genome, and the intensive investigations aimed at understanding the proteome it encodes, has motivated us to initiate studies in mouse glycomics focused on HS. The current study is aimed at determining the quantitative and qualitative organ distribution of HS in mice. HS from brain, eyes, heart, lung, liver, kidney, spleen, intestine and skin was purified from 6-8 week old male and female mice. The recovered yield of HS from these organs is compared with the recovered whole body yield of HS. Structural characterization of the resulting HS relied on disaccharide analysis and 1 H-NMR spectroscopy. Different organs revealed a characteristic HS structure. These data begin to provide a structural understanding of the role of HS in cell-cell interactions, cell signaling

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Section: Structural Characterization Of Organs Hs Using 1 H-nmr Spectmentioning

confidence: 91%

Isolation and characterization of heparan sulfate from various murine tissues

et al. 2006

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“…The recovered solid product (methyl ester of desulfated epimerized chondroitin sulfate) was treated with 10 ml of 0.1 M sodium hydroxide for 24 h at room temperature to obtain free carboxylate, O-desulfated epimerized chondroitin sulfate dCS pII (22). This structure was confirmed by 1 H NMR spectroscopy (23). Desulfated C-5-epimerized Chondroitin Sulfate Trisaccharide (dCS pII3 )-The dCS pII polymer (100 mg) was treated with chondroitin ABC lyase (1 unit) as described earlier.…”

Section: H Nmr and Esi-ms Analysis (Table 3) (21)mentioning

confidence: 94%

Acceptor Specificity of the Pasteurella Hyaluronan and Chondroitin Synthases and Production of Chimeric Glycosaminoglycans

Tracy

Avci

Linhardt

et al. 2007

Journal of Biological Chemistry

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The hyaluronan (HA) synthase, PmHAS, and the chondroitin synthase, PmCS, from the Gram-negative bacterium Pasteurella multocida polymerize the glycosaminoglycan (GAG) sugar chains HA or chondroitin, respectively. The recombinant Escherichia coli-derived enzymes were shown previously to elongate exogenously supplied oligosaccharides of their cognate GAG (e.g. HA elongated by PmHAS). Here we show that oligosaccharides and polysaccharides of certain noncognate GAGs (including sulfated and iduronic acid-containing forms) are elongated by PmHAS (e.g. chondroitin elongated by PmHAS) or PmCS. Various acceptors were tested in assays where the synthase extended the molecule with either a single monosaccharide or a long chain (ϳ10 2-4 sugars). Certain GAGs were very poor acceptors in comparison to the cognate molecules, but elongated products were detected nonetheless. Overall, these findings suggest that for the interaction between the acceptor and the enzyme (a) the orientation of the hydroxyl at the C-4 position of the hexosamine is not critical, (b) the conformation of C-5 of the hexuronic acid (glucuronic versus iduronic) is not crucial, and (c) additional negative sulfate groups are well tolerated in certain cases, such as on C-6 of the hexosamine, but others, including C-4 sulfates, were not or were poorly tolerated. In vivo, the bacterial enzymes only process unsulfated polymers; thus it is not expected that the PmCS and PmHAS catalysts would exhibit such relative relaxed sugar specificity by acting on a variety of animal-derived sulfated or epimerized GAGs. However, this feature allows the chemoenzymatic synthesis of a variety of chimeric GAG polymers, including mimics of proteoglycan complexes. Glycosaminoglycans (GAGs),2 polysaccharides containing a hexosamine as part of their repeat unit, serve essential biological functions in vertebrates, including adhesion, modulation of motility and proliferation, and coagulation (1-8). Certain pathogenic bacteria camouflage themselves with GAGs to increase virulence and enhance infection in their animal hosts. These capsular polysaccharides serve as molecular camouflage as well as a means to potentially hijack host functions (9). The backbones of the vertebrate GAGs, HA (-␤1,4-GlcUA-␤1,3-GlcNAc-), chondroitin sulfate (-␤1,4-GlcUA-␤1,3-GalNAc-), heparan sulfate (␣1,4-GlcUA-␤1,4-GlcNAc-), heparin (-␣1,4-IdoUA-␣ 1,4-GlcNAc-), and keratan sulfate (-␤1,3-Gal-␤1,4-GlcNAc-), are sulfated except for HA. The bacterial GAG polymers are not known to be sulfated. The enzymes involved in the biosynthesis of all the GAG backbones, except for keratan, have been molecularly cloned.Certain bacterial enzymes, including the Gram-negative Pasteurella multocida type A hyaluronan synthase, PmHAS, and the type F chondroitin synthase, PmCS, are particularly amenable to study because of their two active center architecture (10, 11), their ability to polymerize long chains in vitro (12), and their ability to elongate exogenously supplied acceptor oligosaccharides in vitro (13,14). The Escherichia ...

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“…Preparation of desulfated CS-C and CS-D CS-C and CS-D preparations were desulfated as described previously [23]. Briefly, 0.5 mg of each CS preparation was dissolved in 1.5 ml of water and then converted from a sodium form to a hydrogen form using a cation-exchange Dowex 50-X2 column (hydrogen form, 1 ml).…”

Section: Methodsmentioning

confidence: 99%

“…Chemical methods such as acidic hydrolysis [17][18][19][20], methanolysis [21], and hydrazinolysis after deamination [22] have also been used for the characterization of these uronic acid residues, but are associated with a considerable loss of these residues due to the difference in the sensitivity of glycosidic linkages of IdoA and GlcA to conditions for depolymerization and the instability of free uronic acid under neutral/acidic conditions [19,21,23,24]. 1 HNuclear magnetic resonance (NMR) spectroscopy based on the integration of proton signals corresponding to GlcA and IdoA in CS/DS chains has been applied to overcome the difficulties [23], with solvolysis used to remove Osulfate groups to simplify the complex spectrum caused by the variation in sulfation positions. Moreover, a large amount of sample (1-2 mg) is required for the spectroscopy, due to low sensitivity.…”

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confidence: 99%

Determination of iduronic acid and glucuronic acid in sulfated chondroitin/dermatan hybrid chains by 1H-nuclear magnetic resonance spectroscopy

Yamada

Basappa

et al. 2008

Glycoconj J

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The relative proportion of L: -iduronic acid (IdoA) and D: -glucuronic acid (GlcA) is of great importance for the structure-function relationship of chondroitin sulfate (CS)/dermatan sulfate (DS). However, determination of the isotypes of uronic acid residues in CS/DS is still a challenge, due to the instability of free uronic acid released by chemical degradation and its conversion to unsaturated uronic acid by digestion with bacterial eliminase. (1)H-Nuclear magnetic resonance (NMR) spectroscopy is a promising tool with which to address this issue, but the traditional method based on the assignment of the ring proton signals of IdoA and GlcA residues still has drawbacks such as the serious overlap of signals in the (1)H-NMR spectrum of CS/DS polysaccharides. We found that the proton signals of the N-acetyl group of N-acetyl-D: -galactosamines in CS and DS could be clearly distinguished and accurately integrated in the one-dimensional (1D) (1)H-NMR spectrum. Based on this finding, here we report a novel, sensitive, and nondestructive 1D (1)H-NMR-based method to determine the proportion of IdoA and GlcA residues in CS/DS hybrid chains.

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1H Nuclear Magnetic Resonance Spectroscopic Analysis for Determination of Glucuronic and Iduronic Acids in Dermatan Sulfate, Heparin, and Heparan Sulfate

Cited by 36 publications

References 40 publications

Isolation and characterization of heparan sulfate from various murine tissues

Isolation and characterization of heparan sulfate from various murine tissues

Acceptor Specificity of the Pasteurella Hyaluronan and Chondroitin Synthases and Production of Chimeric Glycosaminoglycans

Determination of iduronic acid and glucuronic acid in sulfated chondroitin/dermatan hybrid chains by 1H-nuclear magnetic resonance spectroscopy

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