Patents and literature

Linhardt, Robert J.; Wedlock, D.J.; Phillips, Glyn O.; Balazs, EA; Leshchiner, A.; Langer, Robert S.; Cooney, Courtney; Fitzgerald, George; Grant, Alice H.; Lasker, Sigmund E.; Goulay, J.; Goulay-Heir, M. T.; Goulay-Heir, M. A.; Goulay-Heir, G.; Lormeau, J C; Petitou, Maurice; Choay, J; Soll, D.; Harrison, S. E.; Teng, Li-ong; Walton, A. G.; Sparer, R. V.; Ekwuribe, N.; Bracke, J. W.; Thacker, K.; Brocklehurst, P.; Ferguson, A. S.; Gordon, Lance K.; Hayes, Hélène; Inoue, Sachiko; Yamamoto, Hiroyuki; Nonomura, Arthur M.; Pappo, R.; Schweid, J. M.; Cohee, A. H.; Dec, A. F.; Shay, L. K.; Reiter, Silke; Turrisi, L.; Umezawa, Iwao; Komiyama, Kanki; Williams, Daryl R.; Munnecke, D. M.; Zahradník, R.; Arizono, Koji; Terasawa, Masato; Nobutoki, M.; Borchardt, John K.; Bergwitz, L. C. A.; Osterlund, R. G. L.; Brain, Chelsea M.; Johnston, Blair F.; Brooker, L. G.; Chang, Tien-Hao; Zientek, L. J.; Viniguaz, A.; Chibata, Ichiro; Tosa, Tetsuya; Takamatsu, S.; Doehnert, D. F.; Hill, Alexander S.; Gaehring, D. P.; Scanlon, Jennamarie; Hansen, Gary D.; Hellsten, K. M. E.; Horion, R. L.; Kennedy, Richard B.; Kohn, R. S.; Krüger, Antje; Johnson, Julie K.; Kuu, Wei Y.; Larsson, V. K.; Mezzino, J. F.; Chuang, L. Y.; Michnowski, J.; Miyashiro, Yoshimichi; Yamazaki, Yukiko; Igarasi, Seizi; Morimoto, Kouichi; Nochumson, S.; Pellico, M. A.; Purinton, R. J.; Reinhardt, Frank; Sakata, Kyousuke; Imai, Hiroaki; Sensabaugh, A. J.; Crepps, D. C.; Rice, W. Y.; Fearrington, G. W.; Shim, J. L.; Shimohiro, Y.; Ogawa, Mari; Ohnishi, Tsuyoshi; Simon, Eric J.; Stuhler, H.; Krempl, E.; Su, Donghai; Szendrei, K.; Minker, E.; Re, Z.; Koch, Lori; Wolf, L.; Szijj, E.; Re, M.; Trop, M.; Tunc, D. C.; Ueda, Takuya; Vellini, M.; Rb, Wallace; Watanabe, Kinzo; Yoshikawa, Sadao; Futami, S.; Wunder, R. H.

doi:10.1007/bf02798421

Cited by 2 publications

(2 citation statements)

References 23 publications

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“…The depolymerized freeze-dried samples of chondroitin sulphate and dermatan sulphate were reconstituted at 1 mg/ml. The final UV absorbance value of the enzymic digestion mixture was measured at 232 nm in 30 mM HCl solution (e5500 M-1 * cm-') [6]. The Mr of the disaccharide resulting from the major repeating unit of the polysaccharide chain (see Scheme 1) was used to calculate the mol of product formed.…”

Section: Resultsmentioning

confidence: 99%

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Purification, characterization and specificity of chondroitin lyases and glycuronidase from Flavobacterium heparinum

Linhardt

Laliberte³

et al. 1995

Biochemical Journal

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The chondroitin lyases from Flavobacterium heparinum (Cytophaga heparinia) have been widely used in depolymerization of glycosaminoglycan and proteoglycan chondroitin sulphates. Oligosaccharide products derived from chondroitin sulphate can be further degraded by glycuronidases and sulphatases obtained from the same organism. There has been no reported purification of these enzymes to homogeneity nor is there any information on their physical and kinetic characteristics. The absence of pure enzymes has resulted in a lack of understanding of the optimal conditions for their catalytic activity and their substrate specificity. This has limited the use of these enzymes as reagents for preparation of oligosaccharides for structure and activity studies. Reproducible schemes to purify a chondroitin AC lyase, a glycuronidase and chondroitin B lyase from Flavobacterium heparinum to apparent homogeneity are described. Chondroitin AC lyase (chondroitinase AC, EC 4.2.2.5), glycuronidase [chondro-(1-->3)-glycuronidase, no EC number] and chondroitin B lyase (chondroitinase B, no EC number) have M(r) values (assessed by SDS/PAGE) of 74,000, 41,800 and 55,200 respectively, and isoelectric points (determined by isoelectric focusing) of 8.85, 9.28 and 9.05 respectively. Chondroitin lyase AC and B contain pyroglutamic acid at their N-termini precluding their analysis by Edman degradation. Deblocking with pyroglutamate aminopeptidase facilitated the determination of their N-terminal sequences. The kinetic properties of these enzymes have been determined as well as the optimum conditions for their catalytic activity. The specificity of the glycouronidase, determined using 17 different disaccharide substrates, shows that it only acts on unsulphated or 6-O-sulphated 1-->3 linkages. The chondroitin lyases are both endolytic enzymes, and oligosaccharide mapping shows their expected specificity towards the chondroitin and dermatan sulphate polymers.

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Section: Resultsmentioning

confidence: 99%

“…Two classes of enzyme that act on glycosaminoglycans are polysaccharide lyases and hydrolases. Prokaryotic polysaccharide lyases depolymerize glycosaminoglycans through an eliminative mechanism [6] whereas enzymes from eukaryotic sources act through a hydrolytic mechanism.…”

Section: Introductionmentioning

confidence: 99%

Purification, characterization and specificity of chondroitin lyases and glycuronidase from Flavobacterium heparinum

Linhardt

Laliberte³

et al. 1995

Biochemical Journal

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Ionic Liquid Matrix for Direct UV-MALDI-TOF-MS Analysis of Dermatan Sulfate and Chondroitin Sulfate Oligosaccharides

2006

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Polyanionic oligosaccharides such as dermatan sulfate (DS) and chondroitin sulfate (CS) exhibit poor ionization efficiencies and tend to undergo thermal fragmentation through the loss of SO 3 under conventional ultraviolet matrix-assisted laser desorption/ionization (UV-MALDI) conditions. A new ionic liquid matrix (ILM), a guanidinium salt of α-cyano-4-hydroxycinnamic acid, facilitates direct UV-MALDI mass spectrometric (MS) analysis of underivatized DS and CS oligosaccharides up to a decasaccharide in their common form as sodium salts. The resulting mass spectra show very low extent of fragmentation through an SO 3 loss. The new ILM is suitable for MALDI-MS analysis of mixtures containing oligosaccharides with different numbers of sulfo groups.The use of room-temperature ionic liquids as matrices is a fast growing area of matrixassisted laser desorption/ionization mass spectrometry (MALDI-MS) method development. Ionic liquid matrices (ILMs) have a number of advantages over conventional crystalline matrices including the homogeneity of the analyte-matrix mixture and high vacuum stability of the matrix. 1,2 ILMs have been shown effective in MALDI-MS analysis of several classes of biological molecules including peptides and proteins, 3 phospholipids, 4 oligonucleotides, 5 and neutral oligosaccharides. 6 The goal of this work was to develop a facile MALDI-timeof-flight (TOF)-MS method for the characterization of glycosaminoglycan (GAG)-derived oligosaccharides using an ILM.When analyzed by mass spectrometry, GAG-derived oligosaccharides have a tendency to fragment through the loss of SO 3 (Δ −102 m/z). In electrospray ionization (ESI) mass spectrometry, these polysulfated, polycarboxylated oligosaccharides often result in several peaks, corresponding to intact oligosaccharide and fragments due to the SO 3 loss, 7 each of which can be sodiumcationized, protonated, or both and have various charge states. 8 The advantage of MALDI is that multiply charged species are not observed in the reflector mass spectra, 9 simplifying data interpretation as compared to the ESI mass spectra. However, due to their thermal lability, poor ionization efficiency, and a tendency to undergo extensive Na/H exchange, polyanions such as GAG-derived oligosaccharides are rarely analyzed by MALDI-MS, and methods for such analysis are scarce.To date, a very few methods have been reported for MALDI-MS analysis of underivatized GAG-derived oligosaccharides. 10-12 A peptide complexation method described by Juhasz and Biemann 13,14 allows the detection of polysulfated oligosaccharides by MALDI-MS in a form of noncovalent complexes with basic peptides, (Gly-Arg) n , where n exceeds the number of sulfo groups in the oligosaccharide by one. While this method is sensitive and affords MALDI mass spectra in which SO 3 loss is suppressed, 15 it is limited by the availability of synthetic peptides. Recently, we reported a successful MALDI-MS analysis of sodium salts of an octasulfated disaccharide and an octasulfated pentasaccharide in their uncomp...

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Patents and literature

Cited by 2 publications

References 23 publications

Purification, characterization and specificity of chondroitin lyases and glycuronidase from Flavobacterium heparinum

Purification, characterization and specificity of chondroitin lyases and glycuronidase from Flavobacterium heparinum

Ionic Liquid Matrix for Direct UV-MALDI-TOF-MS Analysis of Dermatan Sulfate and Chondroitin Sulfate Oligosaccharides

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