Solution Structures of Chemoenzymatically Synthesized Heparin and Its Precursors

Zhang, Zhenqing; McCallum, Scott A.; Xie, Jin; Nieto, Lidia; Corzana, Francisco; Jiménez‐Barbero, Jesús; Chen, Miao; Liu, Jian; Linhardt, Robert J.

doi:10.1021/ja8026345

Cited by 149 publications

(163 citation statements)

References 54 publications

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“…Second, the precise structure of the HS oligosaccharide that displays both anti-Xa and anti-IIa activities is unknown. Previously, we demonstrated the synthesis of polysaccharides using HS biosynthetic enzymes and a capsular polysaccharide from the E. coli K5 strain; however, the products are a mixture differing in both the size and distribution of sulfo groups and IdoUA residues, known as sulfation patterns (10,12). In addition, we completed the synthesis of size-and sulfation pattern-defined ULMW heparins; however, the products exhibit only anti-Xa activity because of their short size (14).…”

Section: Discussionmentioning

confidence: 99%

Chemoenzymatic Synthesis of Heparin Oligosaccharides with both Anti-factor Xa and Anti-factor IIa Activities

Pempe

Liu

2012

Journal of Biological Chemistry

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Background: Heparin inhibits the activity of factors Xa and IIa in the blood coagulation cascade. Results: A series of size-defined N-sulfated oligosaccharides were synthesized to probe the size requirement for the oligosaccharides displaying anti-IIa activity. Conclusion: Oligosaccharides that display anti-IIa activity are longer than 19 saccharide residues. Significance: The results will direct efforts to prepare synthetic heparin with both anti-Xa and anti-IIa activities.

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Section: Discussionmentioning

confidence: 99%

Chemoenzymatic Synthesis of Heparin Oligosaccharides with both Anti-factor Xa and Anti-factor IIa Activities

Pempe

Liu

2012

Journal of Biological Chemistry

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“…With the exception of HS polymerase, all of these biosynthetic enzymes have been expressed at high levels in Escherichia coli (1), permitting easy access to an abundance of enzymes. Using HS sulfotransferases and C 5 -epi, we previously developed a method to synthesize HS from a bacteria capsular polysaccharide with biological activities (6,9,10).…”

Section: Heparan Sulfate (Hs)mentioning

confidence: 99%

Chemoenzymatic Design of Heparan Sulfate Oligosaccharides*

Liu

Chen

et al. 2010

Journal of Biological Chemistry

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Heparan sulfate is a sulfated glycan that exhibits essential physiological functions. Interrogation of the specificity of heparan sulfate-mediated activities demands a library of structurally defined oligosaccharides. Chemical synthesis of large heparan sulfate oligosaccharides remains challenging. We report the synthesis of oligosaccharides with different sulfation patterns and sizes from a disaccharide building block using glycosyltransferases, heparan sulfate C 5 -epimerase, and sulfotransferases. This method offers a generic approach to prepare heparan sulfate oligosaccharides possessing predictable structures. Heparan sulfate (HS)3 is a unique class of macromolecular natural product that is present in large quantities on the mammalian cell surface and in the extracellular matrix. HS participates in regulating blood coagulation, embryonic development, and the inflammatory response and assists viral/bacterial infections. It consists of a repeating disaccharide unit of glucuronic acid (GlcUA) or iduronic acid (IdoUA) and glucosamine, both capable of carrying sulfo groups (1). The sulfation pattern of HS dictates its biological activity (2, 3). Heparin, a widely used anticoagulant drug, is a specialized form of highly sulfated HS. The diverse biological functions present considerable opportunities for exploiting HS or HS-protein conjugates for developing new classes of anticancer (4), antiviral (5), and improved anticoagulant drugs (6). Furthermore, a recent worldwide outbreak of contaminated heparin underscores the needs for synthetic heparins to replace those isolated from animal tissues (7). Chemical synthesis is a powerful tool to obtain structurally defined heparin/HS oligosaccharides. The most successful example is the total synthesis of an antithrombin-binding pentasaccharide (8). This pentasaccharide is marketed under the trade name Arixtra for the treatment of venous thromboembolic disorders. However, the chemical synthesis of oligosaccharides larger than an octasaccharide is extremely difficult, especially when multiple target structures are required for biological evaluation (8). An enzyme-based method offers a promising alternative approach to synthesize HS.The HS biosynthetic pathway involves multiple enzymes, including HS polymerase, epimerase, and sulfotransferases ( Fig. 1). HS polymerase is responsible for building the polysaccharide backbone, containing the repeating unit of -GlcUA-GlcNAc-. The backbone is then modified by N-deacetylase/N-sulfotransferase (having two separate domains exhibiting the activity of N-deacetylase and N-sulfotransferase, respectively), C 5 -epimerase (C 5 -epi, converting GlcUA to IdoUA), 2-O-sulfotransferase (2-OST), 6-O-sulfotransferase (6-OST) and 3-O-sulfotransferase (3-OST) to produce the fully elaborated HS. With the exception of HS polymerase, all of these biosynthetic enzymes have been expressed at high levels in Escherichia coli (1), permitting easy access to an abundance of enzymes. Using HS sulfotransferases and C 5 -epi, we previously developed a method ...

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“…We have run the simulation using AMBER 12 27 with the latest GLYCAM parameters 28 and partial charges, 29 which represent a consistent parameterization to model glycosaminoglycans, and it has been employed with that aim. 30 After the appropriate equilibration steps we have run a long simulation (500 ns) in explicit water (TIP3P), 31 periodic boundary conditions (PBC) 32 using the particle mesh Ewald method (PME). 33 Results were consistent with stable 1 C 4 puckering with scarce transitions to 2 S 0 , and a narrow syn-Φ and syn-ψ glycosidic linkage distribution (Fig.…”

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confidence: 99%

3D structure of a heparin mimetic analogue of a FGF-1 activator. A NMR and molecular modelling study

et al. 2013

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The motional behaviour of heparin oligosaccharides in solution is best described as a top rotor having two perpendicular rotation axes. This prevents an accurate extraction of interprotonic distances by NOESY/ROESY based methods. In this paper, we describe the solution structure of the hexasaccharide 1 calculated from high exactitude distance data obtained from off-resonance ROESY combined with a long MD simulation of 500 ns. In previous studies, we have found that two synthetic hexasaccharides having the sulphate groups directed towards one side of its central plane have an opposite biological activity, while 1 is unable to activate the FGF-1 signalling pathway, the other (2) is even more active than the regular region derived hexasaccharide (3) that mimics the natural active compound, heparin. From the structural analysis it was concluded that 1 has similar three-dimensional characteristics to 2 or 3 and therefore the differences in the activity should be due to the arrangement of the sulphate groups within the hexasaccharidic sequence.

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Solution Structures of Chemoenzymatically Synthesized Heparin and Its Precursors

Cited by 149 publications

References 54 publications

Chemoenzymatic Synthesis of Heparin Oligosaccharides with both Anti-factor Xa and Anti-factor IIa Activities

Chemoenzymatic Synthesis of Heparin Oligosaccharides with both Anti-factor Xa and Anti-factor IIa Activities

Chemoenzymatic Design of Heparan Sulfate Oligosaccharides*

3D structure of a heparin mimetic analogue of a FGF-1 activator. A NMR and molecular modelling study

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