Use of biosynthetic enzymes in heparin and heparan sulfate synthesis

Chappell, Elizabeth P.; Liu, Jian

doi:10.1016/j.bmc.2012.11.053

Cited by 38 publications

(23 citation statements)

References 48 publications

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“…HS chains are made up of variably sulfated and N-acetylated repeating disaccharide unites, i.e. glucuronic/iduronic acid and glucosamine (Chappell & Liu, 2013;Linhardt, 2003;Sakiyama-Elbert, 2014). Heparin, the most highly sulfacted heparan sulfate, has been widely used as the most effective clinical anticoagulant.…”

Section: Bioactive Polysaccharides From Other Sourcesmentioning

confidence: 99%

A review of bioactive plant polysaccharides: Biological activities, functionalization, and biomedical applications

Liu

Willför

2015

Bioactive Carbohydrates and Dietary Fibre

506

202

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Section: Bioactive Polysaccharides From Other Sourcesmentioning

confidence: 99%

A review of bioactive plant polysaccharides: Biological activities, functionalization, and biomedical applications

Liu

Willför

2015

Bioactive Carbohydrates and Dietary Fibre

506

202

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“…To generate synthetic heparin with controllable structural and functional profiles, a different combined chemoenzymatic approach was also recently developed Chappell and Liu, 2013;DeAngelis et al, 2013). The enzymes of heparin/HS biosynthesis, expressed in E. coli or insect cells, may be used for postpolymerization transformations of a heparosan-like polymer made using bacterial enzymes.…”

Section: E Synthetic Heparinmentioning

confidence: 99%

Pharmacology of Heparin and Related Drugs

et al. 2015

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“…11,12 Natural isolates are frequently inhomogeneous, making structural characterization difficult. Here, we capitalize on a synthetic strategy directed at the preparation of HS with specific sulfation patterns (Figure.…”

mentioning

confidence: 99%

Structural Aspects of Heparan Sulfate Binding to Robo1–Ig1–2

et al. 2016

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Roundabout 1, or Robo1, is a cell surface signaling molecule important in axon guidance. Its interaction with heparan sulfate (HS) and members of the Slit protein family is essential to its activity, making characterization of these interactions by structural methods, such as NMR, highly desirable. However, the fact that Robo1 is a glycosylated protein prevents employment of commonly used bacterial hosts for expression of properly glycosylated forms with the uniform 15N, 13C, and 2H labeling needed for NMR studies. Here, we apply an alternative methodology, based on labeling with a single amino acid type and high structural content NMR data, to characterize a two-domain construct of glycosylated Robo1 (Robo1–Ig1–2) interacting with a synthetic HS tetramer (IdoA-GlcNS6S-IdoA2S-GlcNS6S-(CH2)5NH2). Significant chemical shift perturbations of the crosspeak from K81 on titration with the tetramer provide initial evidence for the location of a binding site and allow determination of a 255 μM disassociation constant. The binding epitopes, bound conformation, and binding site placement of the HS tetramer have been further characterized by saturation transfer difference (STD), transferred nuclear Overhauser effect (trNOE), and paramagnetic perturbation experiments. A model of the complex has been generated using constraints derived from the various NMR experiments. Postprocessing energetic analysis of this model provides a rationale for the role each glycan residue plays in the binding event, and examination of the binding site in the context of a previous Robo-Slit structure provides a rationale for modulation of Robo-Slit interactions by HS.

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Use of biosynthetic enzymes in heparin and heparan sulfate synthesis

Cited by 38 publications

References 48 publications

A review of bioactive plant polysaccharides: Biological activities, functionalization, and biomedical applications

A review of bioactive plant polysaccharides: Biological activities, functionalization, and biomedical applications

Pharmacology of Heparin and Related Drugs

Structural Aspects of Heparan Sulfate Binding to Robo1–Ig1–2

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