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

Xu, Yongmei; Pempe, Elizabeth H.; Liu, Jian

doi:10.1074/jbc.m112.358523

Cited by 55 publications

(59 citation statements)

References 31 publications

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“…New “top-down” analytical methods based on tandem mass spectrometry and ion-mobility methods hold great promise for providing much greater contextual information about the structures of 3- O -sulfation sites (Kailemia et al, 2013; Meissen et al, 2009). Chemical and chemoenzymatic methods have been recently developed that have the capacity to produce multimilligram quantities of defined oligosaccharides, providing much needed reagents to aid in the discovery of new ligands and to characterize the binding sites in the enzymes and in the protein ligands (Arungundram et al, 2009; Xu et al, 2011; Xu et al, 2012). Genetic studies of model organisms focused on 3- O -sulfation have begun to yield interesting insights into the biological function of the Hs3sts and new ligands to study.…”

Section: Discussionmentioning

confidence: 99%

Heparan sulfate 3-O-sulfation: A rare modification in search of a function

Thacker¹,

Xu²,

Lawrence³

et al. 2014

Matrix Biology

199

244

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Many protein ligands bind to heparan sulfate, which results in their presentation, protection, oligomerization or conformational activation. Binding depends on the pattern of sulfation and arrangement of uronic acid epimers along the chains. Sulfation at the C3 position of glucosamine is a relatively rare, yet biologically significant modification, initially described as a key determinant for binding and activation of antithrombin and later for infection by Type I Herpes Simplex virus. In mammals, a family of seven heparan sulfate 3-O-sulfotransferases installs sulfate groups at this position and constitutes the largest group of sulfotransferases involved in heparan sulfate formation. However, to date very few proteins or biological systems have been described that are influenced by 3-O-sulfation. This review describes our current understanding of the prevalence and structure of 3-O-sulfation sites, expression and substrate specificity of the 3-O-sulfotransferase family and the emerging roles of 3-O-sulfation in biology.

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Section: Discussionmentioning

confidence: 99%

Heparan sulfate 3-O-sulfation: A rare modification in search of a function

Thacker¹,

Xu²,

Lawrence³

et al. 2014

Matrix Biology

199

244

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“…The expression of these enzymes were carried out in Escherichia coli and purified by an amylose-agarose column (New England Biolabs) as previously described (53).…”

Section: Methodsmentioning

confidence: 99%

Fibroblast Growth Factor-based Signaling through Synthetic Heparan Sulfate Blocks Copolymers Studied Using High Cell Density Three-dimensional Cell Printing

Sterner

Masuko

et al. 2014

Journal of Biological Chemistry

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Background: Fibroblast growth factor-receptor-heparan sulfate (FGF-HS-FGFR) signals cell proliferation. Results: HS synthesized with sulfated domains at its non-reducing ends actively promotes cellular proliferation in a threedimensional cell microarray. Conclusion: A symmetric 2:2:2 FGF-HS-FGFR complex is preferred over an asymmetric 2:1:2 model by these data. Significance: This paper suggests a preference for symmetry in the signal transduction complex having two FGF-FGFR on the non-reducing end of two HS chains.

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“…The following enzymes and appropriate expression vectors and bacterial expression cells, prepared as previously described [26]. These enzymes include rat AST-IV (EC 2.8.2.1) in pET15 expression vector and BL21 expression cells; human C 5 -epimerase (NCBI: NM_015554.1), mouse 6 O ST isoform 1 (NCBI: NM_015818.2), and mouse 6 O ST isoform 3 (NCBI: NM_015820.3) in pMAL-c2x expression vector and Rosetta-gami-B with GroEL expression cells; hamster 2- O ST (GenBank: D88811.1) in pMAL-c2x expression vector and Rosetta-gami-B expression cells; mouse 3 O ST-1 (NCBI: NM_010474.2) in pET28 expression vector and BL21(DE3)RIL expression cells.…”

Section: Methodsmentioning

confidence: 99%

Assays for determining heparan sulfate and heparin O-sulfotransferase activity and specificity

Sterner

Paul

et al. 2013

Anal Bioanal Chem

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O-sulfotransferases (OSTs) are critical enzymes in the cellular biosynthesis of the biologically and pharmacologically important heparan sulfate and heparin. Recently, these enzymes have been cloned and expressed in bacteria for application in the chemoenzymatic synthesis of glycosaminoglycan-based drugs. OST activity assays have largely relied on the use of radioisotopic methods using [35S] 3'-phosphoadenosine-5'-phosphosulfate and scintillation counting. Herein, we examine alternative assays that are more compatible with a biomanufacturing environment. A high throughput microtiter-based approach is reported that relies on a coupled bienzymic colorimetric assay for heparan sulfate and heparin OSTs acting on polysaccharide substrates using arylsulfotransferase-IV and p-nitrophenylsulfate as a sacrificial sulfogroup donor. A second liquid chromatography-mass spectrometric assay, for heparan sulfate and heparin OSTs acting on structurally defined oligosaccharide substrates, is also reported that provides additional information on the number and positions of the transferred sulfo groups within the product. Together, these assays allow quantitative and mechanistic information to be obtained on OSTs that act on heparan sulfate and heparin precursors.

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

Cited by 55 publications

References 31 publications

Heparan sulfate 3-O-sulfation: A rare modification in search of a function

Heparan sulfate 3-O-sulfation: A rare modification in search of a function

Fibroblast Growth Factor-based Signaling through Synthetic Heparan Sulfate Blocks Copolymers Studied Using High Cell Density Three-dimensional Cell Printing

Assays for determining heparan sulfate and heparin O-sulfotransferase activity and specificity

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