Chondroitin O-methyl ester: an unusual substrate for chondroitin AC lyase

Avci, Fikri Y.; Toida, Toshihiko; Linhardt, Robert J.

doi:10.1016/s0008-6215(03)00348-3

Cited by 7 publications

(12 citation statements)

References 16 publications

(12 reference statements)

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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).…”

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

confidence: 93%

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

confidence: 93%

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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“…PVA and several polysaccharides have been modified with acrylates and methacrylates by different ways [17][18][19][20]. The addition of unsaturations to polymers allows the reticulation of modified polymers without the addition of cross-linking agents [21][22][23][24][25][26]. The reaction may be carried out through the radical initiator pathway or by UV-light [27].…”

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confidence: 99%

Hydrogels based on chemically modified poly(vinyl alcohol) (PVA-GMA) and PVA-GMA/chondroitin sulfate: Preparation and characterization

Crispim¹,

Piai²,

Fajardo³

et al. 2012

Express Polym. Lett.

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This work reports the preparation of hydrogels based on PVA-GMA, PVA-GMA is poly(vinyl alcohol) (PVA) functionalized with vinyl groups from glycidyl methacrylate (GMA), and on PVA-GMA with different content of chondroitin sulfate (CS). The degrees of swelling of PVA-GMA and PVA-GMA/CS hydrogels were evaluated in distilled water and the swelling kinetics was performed in simulated gastric and intestinal fluids (SGF and SIF). PVA-GMA and PVAGMA/CS hydrogels demonstrated to be resistant on SGF and SIF fluids. The elastic modulus, E, of swollen-hydrogels were determined through compressive tests and, according to the obtained results, the hydrogels presented good mechanical properties. At last, the presence of CS enhances the hydrogel cell compatibility as gathered by cytotoxicity assays. It was concluded that the hydrogels prepared through this work presented characteristics that allow them to be used as biomaterial, as a carrier in drug delivery system or to act as scaffolds in tissue engineering as well

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“…Chondroitin lyase ACI Flavo and chondroitin lyase ACII Arthro both afforded a single major product from hyaluronan migrating at 7.5 min and a different major product migrating at 5.2 min when using the methyl ester of hyaluronan as a substrate. Since methyl-esterification of a carboxy group at the nonreducing end decreases the absorbtivity coefficient of the product compared to the product from intact hyaluronan at 230 nm as indicated previously, 33 the response on CE profile of each product from methylesterified hyaluronan was smaller than those of the products from intact hyaluronan. The migration velocity of molecules on normal polarity is ordered as those of cationic, neutral, and anionic molecules.…”

Section: Enzymatic Degradation Of the Methyl Ester Of Hyaluronan And mentioning

confidence: 56%

“…33 This is in contrast to the formation of only a disaccharide from treatment of methyl ester of hyaluronan with chondroitin lyase ACI Flavo. Furthermore, chondroitin lyase ACII Arthro showed no observable activities on the methyl ester of chondroitin, whereas it showed substantial activity on methyl-esterified hyaluronan.…”

Section: Enzymatic Degradation Of the Methyl Ester Of Hyaluronan And mentioning

confidence: 96%

“…While the action of chondroitin lyase ACI Flavo on the methyl ester of chondroitin has been previously reported, chondroitin lyase ACII Arthro was not observed to act on this substrate. 33 The discovery of this new substrate, the methyl ester of hyaluronan, and the activity of both chondroitin lyase ACI Flavo and chondroitin lyase ACII Arthro is significant in that it demonstrates that the negatively charged D-glucopyranosyluronic acid residue is not required by either enzyme, thus, further clarifying its role on the activity of both chondroitin lyases.…”

Section: Introductionmentioning

confidence: 99%

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Preparation of the methyl ester of hyaluronan and its enzymatic degradation

Hirano

Sakai

Ishikawa

et al. 2005

Carbohydrate Research

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A methyl ester of hyaluronan in which the carboxyl groups were fully esterified was prepared using trimethylsilyl diazomethane. This derivative, while not depolymerized by hyaluronan lyases or hyaluronan hydrolases, was a substrate for both chondroitin ACI lyase (EC 4.2.2.5) from Flavobacterium heparinum and chondroitin ACII lyase (EC 4.2.2.5) from Arthrobacter aurescens. The major product isolated in these depolymerization reactions was methyl α-L-threo-hex-4-enepyranosyluronate-(1→3)-2-acetamido-2-deoxy-α,β-D-glucopyranoside as determined by 1 H NMR spectroscopy and MALDITOF mass spectrometry.

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Chondroitin O-methyl ester: an unusual substrate for chondroitin AC lyase

Cited by 7 publications

References 16 publications

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

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

Hydrogels based on chemically modified poly(vinyl alcohol) (PVA-GMA) and PVA-GMA/chondroitin sulfate: Preparation and characterization

Preparation of the methyl ester of hyaluronan and its enzymatic degradation

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