Electrophoretic analysis of di- and oligosaccharides derived from glycosaminoglycans on microchip format

Matsuno, Yu-ki; Kinoshita, Mitsuhiro; Kakehi, Kazuaki

doi:10.1016/j.jpba.2004.05.020

Cited by 13 publications

(8 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Size-fractionated oligosaccharide K was the first peak eluted out in the Sephadex P-4 chromatography, and thus fraction K contained the largest oligosaccharides in the oligosaccharides (1000 Da , M r , 8000 Da). In addition, larger oligosaccharides were observed earlier based on charge/molecular sizes in CE [48]. Component X was the first peak migrated out in CE.…”

Section: Discussionmentioning

confidence: 62%

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry

et al. 2005

View full text Add to dashboard Cite

A capillary electrophoresis (CE) method was developed for the separation of heparin oligosaccharides compatible to study the interactions between the oligosaccharides and granulocyte-colony stimulating factor (G-CSF). Unfractionated heparin was eliminitively degraded to heparin oligosaccharides by an endolytic heparinase. The degraded smaller oligosaccharides (M(r) < 1000) were baseline-separated by CE under a 50 mM phosphate buffer (pH 9.0) in 10 min. Standard heparin disaccharides and larger oligosaccharides (1000 < M(r) < 8000) were all separated under optimized separation conditions. Compared with standard heparin disaccharides, smaller oligosaccharides contained one nonsulfated, two monosulfated, and two disulfated disaccharides, but trisulfated disaccharides were not found. The smaller oligosaccharides were also identified and molecular mass was deduced by electrospray ionization-mass spectrometry (ESI-MS). Furthermore, interactions between G-CSF and the oligosaccharides were studied by using capillary zone electrophoresis (CZE) under the above separation conditions. It was found that larger oligosaccharides could interact with G-CSF while smaller oligosaccharides were not observed to bind to G-CSF under the experimental conditions. In conclusion, the purified heparinase could selectively degrade heparin into oligosaccharides and the interaction between G-CSF and heparin was correlated with the chain length of heparin.

show abstract

Section: Discussionmentioning

confidence: 62%

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry

et al. 2005

View full text Add to dashboard Cite

show abstract

“…The undigested materials on the filter of Microcon YM-3 were recovered in 0.1 M ammonium acetate, pH 7.0, containing 0.01% bovine serum albumin, and digested at 37°C for 8 h with both heparitinase I and II (0.03 units/ml each); the HS lyase products were recovered with Microcon YM-3. The recovered samples were dried and then fluorotagged at 90°C for 30 min with 2-aminoacridone hydrochloride (27). The fluorotagged disaccharide units of HA, CS, and DS were immediately separated on separating gels (19.5% acrylamide, 0.52% N,NЈ-methylenebisacrylamide, 2.5% glycerol, 0.05% ammonium persulfate, 0.6% agarose, 0.1% TEMED, and 0.045 M Tris acetate buffer, pH 7.0) with a stacking gel (7.5% acrylamide, 0.2% N,NЈ-methylenebisacrylamide, 2.5% glycerol, 0.05% ammonium persulfate, 0.6% agarose, 0.1% TEMED, and 0.045 M Tris acetate buffer, pH 7.0).…”

Section: 1) Gags (Chondroitin Sulfate a (Cs-mentioning

confidence: 99%

Identification and Functions of Chondroitin Sulfate in the Milieu of Neural Stem Cells

Ida

Shuo

Hirano

et al. 2006

Journal of Biological Chemistry

126

105

View full text Add to dashboard Cite

The behavior of cells is generally considered to be regulated by environmental factors, but the molecules in the milieu of neural stem cells have been little studied. We found by immunohistochemistry that chondroitin sulfate (CS) existed in the surroundings of nestin-positive cells or neural stem/progenitor cells in the rat ventricular zone of the telencephalon at embryonic day 14. Brain-specific chondroitin sulfate proteoglycans (CSPGs), including neurocan, phosphacan/receptor-type protein-tyrosine phosphatase ␤, and neuroglycan C, were detected in the ventricular zone. Neurospheres formed by cells from the fetal telencephalon also expressed these CSPGs and NG2 proteoglycan. To examine the structural features and functions of CS polysaccharides in the milieu of neural stem cells, we isolated and purified CS from embryonic day 14 telencephalons. The CS preparation consisted of two fractions differing in size and extent of sulfation: small CS polysaccharides with low sulfation and large CS polysaccharides with high sulfation. Interestingly, both CS polysaccharides and commercial preparations of dermatan sulfate CS-B and an E-type of highly sulfated CS promoted the fibroblast growth factor-2-mediated proliferation of neural stem/progenitor cells. None of these CS preparations promoted the epidermal growth factor-mediated neural stem cell proliferation. These results suggest that these CSPGs are involved in the proliferation of neural stem cells as a group of cell microenvironmental factors.

show abstract

“…Fluorescent labeling of unsaturated disaccharides with AMAC was performed according to the previously reported procedure [13]. HA samples (100 mg) were dissolved in a 20 mM Tri-HCl buffer (pH 8.0, 100 ml) and an aqueous solution of chondroitinase ABC (100 mU/2 ml) was added, and the mixture was incubated at 371C for 16 h. After keeping the mixture on the boiling water bath for 5 min, the mixture was centrifuged at 8000 Â g for 10 min and the supernatant was lyophilized to dryness.…”

Section: Fluorescent Labeling Of Unsaturated Disaccharides Derived Frmentioning

confidence: 99%

Electrophoresis studies on the contaminating glycosaminoglycans in commercially available hyaluronic acid products

et al. 2008

Self Cite

View full text Add to dashboard Cite

Hyaluronic acid (HA) samples showing inhibition effect on digestion with testicular hyaluronidase (HAase) were found from 16 commercially available HA products, which were supplied from 11 different manufacturers. Most of these HA samples (six samples) were derived from the rooster comb, and one sample was derived from the human umbilical cord. HA oligosaccharides produced by exhaustive digestion of these HA samples with testicular HAases were monitored by capillary electrophoresis, and we found that a few HA samples gave no oligosaccharide products. Detailed analysis of HA samples by cellulose acetate membrane electrophoresis revealed that the HA samples were not digested with HAase because of the presence of a small amount of dermatan sulfate (DS). Analysis of disaccharide units of these HA samples produced by digestion with chondroitinase ABC supported the observations. And the content of DS in the sample was estimated to be ca. 8%. In contrast, these HA samples were easily digested with bacterial hyaluronate lyases from Streptomyces hyalurolyticus and Streptococcus dysgalactiae and gave endproducts of unsaturated disaccharide or unsaturated tetra- or hexasaccharides. The results suggested that the inhibitory effect of DS on HAase is specific to endo-type hydrolase (i.e. testicular HAase). In addition, pharmaceutical preparations of HA derived from rooster comb were easily digested with testicular HAase. These findings will be useful information for clinical or cosmetic use of HA preparations in terms of their half-life.

show abstract

Electrophoretic analysis of di- and oligosaccharides derived from glycosaminoglycans on microchip format

Cited by 13 publications

References 23 publications

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry

Separation, identification, and interaction of heparin oligosaccharides with granulocyte-colony stimulating factor using capillary electrophoresis and mass spectrometry

Identification and Functions of Chondroitin Sulfate in the Milieu of Neural Stem Cells

Electrophoresis studies on the contaminating glycosaminoglycans in commercially available hyaluronic acid products

Contact Info

Product

Resources

About