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  • Depolymerization of Heparin/HS

    Heparin and heparan sulfate (HS) glycosaminoglycans are linear sulfated polysaccharides located on cell-surface membranes and in extracellular matrices in virtually all animal tissues. Heparin and HS have been implicated in cell-biological processes, cell adhesion and regulation of enzymatic catalysis (1). HS chains have been shown to interact with a variety of growth factors, chemokines, extracellular matrix proteins, and enzymes, including antithrombin, fibroblast growth factors, and vascular endothelial growth factor (2) as shown by choosing the tab below. Heparin has been widely used as an anticoagulant drug (3,4), and it has been shown to regulate cellular process by binding, stabilizing and activating various growth factors (5).

    References

    1. Fritz, T., et al (1994) J Biol Chem 269(46), 28809-28814. PMID: 7961837
    2. Linhardt, R. J., et al. (1990) Biochemistry 29(10), 2611-2617. PMID: 2334685
    3. Linhardt, R. J. and Gunay, N. S. (1999) Semin. Thromb. Hemost. 25 Suppl. 3, 5-16. PMID: 10549711
    4. Casu, B., et al. (2002) Biochemistry 41(33), 10519-10528. PMID: 12173939
    5. Knudsen, C. B., Knudsen, W. (2001) Semin. Cell. Dev. Biol. 12, 69-78. PMID: 11292372
    6. Ilan N, Elkin M, Vlodavsky I. (2006) Int. J. Biochem. Cell. Biol. 38(12), 2018-39. PMID: 16901744
    7. Rosen SD, Lemjabbar-Alaoui H. (2010) Expert Opin. Ther. Targets. 14(9), 935-49. PMID: 20629619
    • Elucidating the Complexity of Heparin Oligosaccharide Analysis

      Learn more about the structural elucidation of complex and diverse heparin oligosaccharides using Bacteroides Heparinase I, II and III in combination with downstream mass spec analysis.

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    • Overview of Glycobiology

      Learn about the core sequences and common modifications of N-linked and O-linked glycans in this video. Analysis of these glycans and/or peptide portions of the glycoprotein can be accomplished with the use of deglycosylation enzymes, which are explained in detail. Unlike other chemical deglycosylation methods, enzymatic treatment is much gentler and can provide complete sugar removal with no protein degradation.

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    • Identification and Characterization of Protein Glycosylation

      Here we illustrate the use of glycosidases for the analysis of a model glycoprotein: recombinant human chorionic gonadotropin beta (hCGβ), which carries both N-glycans and O-glycans in this video. The technique requires only simple instrumentation and typical consumables, and it can be readily adapted to the analysis of multiple glycoprotein samples.

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    Depolymerization of Heparin/HS includes these areas of focus:

    MS Analysis of GAGs

    HS chain interactions

    Membrane bound proteoglycan (syndecan) showing heparan sulfate chains binding to vascular endothelial growth factor (VEGF). Highly sulfated domains bind free VEGF, creating a morphogen gradient, and thus facilitating binding to its receptor in the plasma membrane. Heparin-bound VEGF is released upon desulfation by sulfatases. Alternatively, active heparin fragments are shed by heparinases. These mechanisms are critical during blood vessel morphogenesis, particularly in tumor proliferation (6,7).