Antithrombin-binding Octasaccharides and Role of Extensions of the Active Pentasaccharide Sequence in the Specificity and Strength of Interaction

Guerrini, Marco; Guglieri, Sara; Casu, Benito; Torri, Giangiacomo; Mourier, Pierre; Boudier, Christian; Viskov, Christian

doi:10.1074/jbc.m801102200

Cited by 78 publications

(80 citation statements)

References 39 publications

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“…ΔU 2S H NS,6S G 2S H NS (α) (peak 9), ΔU 2S H NS,6S G 2S H NS,6S (α), and ΔU 2S H NS,6S G 2S H NS,6S (β) (peaks 14 and 15) are representative of the G 2S -containing species. NMR time course studies conducted for monitoring the depolymerization of heparin benzyl ester demonstrated that the formation of G 2S occurs during the depolymerization process, possibly due to the epimerization of the I 2S -benzyl ester at the C5 position [28]. Table 6 Relative percentage of G 2S , GalA, and epoxide compared to the total uronic acid content at different reaction time points Changes in the precipitation conditions used to isolate enoxaparin result in preferential recovery of hexasulfated tetrasaccharides (peaks 12-15) and loss of undersulfated materials (peaks 1-11).…”

Section: Discussionmentioning

confidence: 99%

Structural elucidation of the tetrasaccharide pool in enoxaparin sodium

et al. 2012

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Low-molecular-weight heparins (LMWHs) are produced from heparin by various depolymerization strategies, which result in a reduction of the average molecular weight of the polysaccharide chains, a reduction of the anti-factor IIa activity (and a concomitant increase in the anti-factor Xa/anti-factor IIa ratio), and introduction of process-related structural signatures. Numerous techniques have been developed to characterize LMWHs and to measure the type and extent of structural modifications that are introduced as a function of the depolymerization process. We present here an analysis of the tetrasaccharide pool of enoxaparin sodium, a LMWH produced by chemical β-elimination of heparin benzyl ester. We identify the predominant sequences present within the tetrasaccharide pool and demonstrate that this pool provides a sensitive, specific readout of the physicochemical process conditions used to generate enoxaparin sodium.

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Section: Discussionmentioning

confidence: 99%

Structural elucidation of the tetrasaccharide pool in enoxaparin sodium

et al. 2012

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“…In addition, both octasaccharides showed a small STD effect for the N-acetyl signal at 2.08 ppm, indicating that this group is oriented toward the opposite side of the AT surface, as observed in all previously studied octasaccharides. 19 No particular differences between STD profiles of Octa-5 and Octa-6 were observed. The overall reduction of the STD intensities (about 15-20%) observed for Octa-6 with respect to Octa-5 is due to a slightly different protein-ligand ratio of the two octasaccharide/AT complexes.…”

Section: Articlementioning

confidence: 90%

Effects on Molecular Conformation and Anticoagulant Activities of 1,6-Anhydrosugars at the Reducing Terminal of Antithrombin-Binding Octasaccharides Isolated from Low-Molecular-Weight Heparin Enoxaparin

et al. 2010

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Terminal 1,6-anhydro-aminosugars (1,6-anAS) are typical structural moieties of enoxaparin, a low-molecular-weight heparin (LMWH) widely used for prevention and treatment of thrombotic disorders. In the enoxaparin manufacturing process, these modified amino sugars are formed during the β-eliminative cleavage of heparin. To investigate the effect of terminal anAS on antithrombin (AT) binding and on inhibition of factor Xa (FXa), two octasaccharides containing modified AT-binding pentasaccharide sequences were isolated from enoxaparin. The molecular conformation of the octasaccharides terminating with N-sulfo-1,6-anhydro-D-mannosamine and N-sulfo-1,6-anhydro-D-glucosamine, respectively, has been determined both in the absence and presence of AT by NMR experiments and docking simulations. Reduced overall contacts of the terminal anAS residues with the binding region of AT induce a decrease in affinity for AT as well as lower anti-FXa activity. The anti-FXa measured either in buffer or plasma milieu does not show any significant difference, suggesting that the inhibition of anti-FXa remains specific and biologically relevant.

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“…HS or heparin forms a 1:1 complex with AT, which deactivates the activities of factor Xa and IIa in the blood coagulation cascade, to exhibit the anticoagulant activity. Anticoagulant HS and heparin isolated from natural sources have an AT-binding pentasaccharide with a structure of -GlcNAc6S-GlcA-GlcNS3SϮ6S-IdoA2S-GlcNS6S-(where GlcNS3Sϩ6S represents N-sulfo glucosamine 3-O-sulfate with or without 6-O-sulfate) (19,20). The IdoA2S residue in the pentasaccharide domain is known to be essential for high ATbinding affinity (21).…”

Section: Resultsmentioning

confidence: 99%

Uncovering Biphasic Catalytic Mode of C5-epimerase in Heparan Sulfate Biosynthesis

Sheng

Dulaney

et al. 2012

Journal of Biological Chemistry

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Background: C 5 -epimerase converts a glucuronic acid to an iduronic acid residue in the heparan sulfate biosynthetic pathway. Results: C 5 -epimerase displays both "reversible" and "irreversible" catalytic modes. Conclusion: C 5 -epimerase recognizes the saccharide sequence of the substrate to position the iduronic acid. Significance: The biphasic catalytic mode of C 5 -epimerase reveals a unique control mechanism in the biosynthesis of heparan sulfate.

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Antithrombin-binding Octasaccharides and Role of Extensions of the Active Pentasaccharide Sequence in the Specificity and Strength of Interaction

Cited by 78 publications

References 39 publications

Structural elucidation of the tetrasaccharide pool in enoxaparin sodium

Structural elucidation of the tetrasaccharide pool in enoxaparin sodium

Effects on Molecular Conformation and Anticoagulant Activities of 1,6-Anhydrosugars at the Reducing Terminal of Antithrombin-Binding Octasaccharides Isolated from Low-Molecular-Weight Heparin Enoxaparin

Uncovering Biphasic Catalytic Mode of C5-epimerase in Heparan Sulfate Biosynthesis

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