Separation of Glycosaminoglycan-Derived Oligosaccharides by Capillary Electrophoresis Using Reverse Polarity

Pervin, Azra; Al-Hakim, Ali; Linhardt, Robert J.

doi:10.1006/abio.1994.1395

Cited by 116 publications

(67 citation statements)

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“…The sample was separated and analyzed using a freshly prepared fused silica capillary (75 m i.d., 375 m o.d., 65 cm long). Reverse polarity analyses were carried out for 25 min at 18 kV using 20 mM phosphoric acid buffer adjusted to pH 3.48 with a saturated solution of dibasic sodium phosphate (25).…”

Section: Purification Of Sized Oligosaccharides Into Homogeneous Oligmentioning

confidence: 99%

“…These efforts were complicated by both the unavailability of pure heparin lyases (22) and the ability of these lyases to cleave glycosidic linkages that are found within the ATIII binding site (14,(31)(32)(33). The heparin lyase sensitivity of glycosidic linkages, within the ATIII binding site, can be overcome, in part, by performing a controlled partial depolymerization (24,25,41). Ototani and Yosizawa (39) used this approach to isolate an octasaccharide with ATIII affinity and anticoagulant activity using heparinase.…”

Section: Enzymatic Preparation Of Antithrombin Binding Sitesmentioning

confidence: 99%

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Enzymatic Preparation of Heparin Oligosaccharides Containing Antithrombin III Binding Sites

Toida

Hileman

Smith

et al. 1996

Journal of Biological Chemistry

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Two new oligosaccharides were prepared from heparin by its partial depolymerization using heparin lyase I (EC 4.2.2.7) in an attempt to prepare oligosaccharides having intact antithrombin III binding sites. The oligosaccharides were purified by chromatography on the basis of both size and charge and demonstrated a high level of purity by capillary electrophoresis. One-and two-dimensional 1 H NMR spectroscopy at 500 MHz revealed the structure of each oligosaccharide. The octasaccharide and decasaccharide are ⌬UAp2S (134) The octasaccharide and decasaccharide correspond to the predominant structural motif found in porcine intestinal mucosal heparin. Sufficient quantities of the decasaccharide were obtained to examine its interaction with antithrombin III using microtitration calorimetry. This decasaccharide bound to antithrombin III with similar avidity as heparin and showed comparable anticoagulant activity, as determined using an antithrombin III dependent anti-factor Xa assay. Interestingly, while both decasaccharide and heparin bound to antithrombin with nanomolar affinity, very little heat of binding was observed.

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Section: Purification Of Sized Oligosaccharides Into Homogeneous Oligmentioning

confidence: 99%

Section: Enzymatic Preparation Of Antithrombin Binding Sitesmentioning

confidence: 99%

Enzymatic Preparation of Heparin Oligosaccharides Containing Antithrombin III Binding Sites

Toida

Hileman

Smith

et al. 1996

Journal of Biological Chemistry

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“…The CE system was operated in the reverse polarity mode by applying the sample at the cathode and run with 20 mM phosphoric acid adjusted to pH 3.5 with 1 M dibasic sodium phosphate as described previously (18). The capillary (75-m inner diameter, 375-m outer diameter, 68 cm long) was manually washed before use with 0.5 ml of 0.5 M sodium hydroxide followed by 0.5 ml of distilled water and then 0.5 ml running buffer.…”

Section: Analysis Of Oligosaccharides Formed From Lyase Treatment Of mentioning

confidence: 99%

A New Glycosaminoglycan from the Giant African Snail Achatina fulica

Kim

Chang

et al. 1996

Journal of Biological Chemistry

118

108

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A new glycosaminoglycan has been isolated from the giant African snail Achatina fulica. This polysaccharide had a molecular weight of 29,000, calculated based on the viscometry, and a uniform repeating disaccharide structure of 34)-2-acetyl,2-deoxy-␣-D-glucopyranose (134)-2-sulfo-␣-L-idopyranosyluronic acid (13. This polysaccharide represents a new, previously undescribed glycosaminoglycan. It is related to the heparin and heparan sulfate families of glycosaminoglycans but is distinctly different from all known members of these classes of glycosaminoglycans. The structure of this polysaccharide, with adjacent N-acetylglucosamine and 2-sulfo-iduronic acid residues, also poses interesting questions about how it is made in light of our current understanding of the biosynthesis of heparin and heparan sulfate. This glycosaminoglycan represents 3-5% of the dry weight of this snail's soft body tissues, suggesting important biological roles for the survival of this organism, and may offer new means to control this pest. Snail glycosaminoglycan tightly binds divalent cations, such as copper(II), suggesting a primary role in metal uptake in the snail. Finally, this new polysaccharide might be applied, like the Escherichia coli K5 capsular polysaccharide, to the study of glycosaminoglycan biosynthesis and to the semisynthesis of new glycosaminoglycan analogs having important biological activities.

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“…Current approaches involve the preparation of CS/DS oligosaccharides using chondroitin lyases followed by separation techniques including gel permeation chromatography (GPC) (Yang et al, 2000), strong anion exhangehigh performance liquid chromatography (SAX-HPLC) Yang et al, 2000), polyacrylamide gel electrophoresis (PAGE) , and capillary electrophoresis (CE) Pervin et al, 1993Pervin et al, , 1994. These provide important data on composition and domain structure but generally yield indirect and incomplete sequence information.…”

Section: Analytical Applicationsmentioning

confidence: 99%

CS Lyases: Structure, Activity, and Applications in Analysis and the Treatment of Diseases

Linhardt

Avci

Toida

et al. 2006

Advances in Pharmacology

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Separation of Glycosaminoglycan-Derived Oligosaccharides by Capillary Electrophoresis Using Reverse Polarity

Cited by 116 publications

References 0 publications

Enzymatic Preparation of Heparin Oligosaccharides Containing Antithrombin III Binding Sites

Enzymatic Preparation of Heparin Oligosaccharides Containing Antithrombin III Binding Sites

A New Glycosaminoglycan from the Giant African Snail Achatina fulica

CS Lyases: Structure, Activity, and Applications in Analysis and the Treatment of Diseases

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