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Enzymatic Sequencing of Heparin Oligosaccharides Using Exolyase

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Proteoglycans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2619))

Abstract

Heparin/heparan sulfate (HP/HS) is a class of acidic polysaccharides with many potential medical applications, especially HP, and its derivatives, low molecular weight heparins (LMWHs), have been widely used as anticoagulants to treat thrombosis for decades. However, the complex structure endows HP/HS a variety of biological functions and hinders the structural and functional studies of HP/HS. Heparinases derived from bacteria are useful tools for the structural studies of HP/HS as well as the preparation of LMWHs. The enzymatic method for the structural analysis of HP/HS chains is easy to operate, requires less samples, and is low cost. Here, we describe an enzymatic approach to investigate the primary sequences of the HP/HS oligosaccharides using a recently discovered exotype heparinase.

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References

  1. Linhardt RJ (2003) 2003 Claude S. Hudson Award address in carbohydrate chemistry Heparin: structure and activity. J Med Chem 46:2551–2564

    Article  CAS  Google Scholar 

  2. Laremore TN, Zhang F, Dordick JS et al (2009) Recent progress and applications in glycosaminoglycan and heparin research. Curr Opin Chem Biol 13:633–660

    Article  CAS  Google Scholar 

  3. Boyce A, Walsh G (2022) Production, characteristics and applications of microbial heparinases. Biochimie 198:109–140

    Article  CAS  Google Scholar 

  4. Zhang Q, Cao HY, Wei L et al (2021) Discovery of exolytic heparinases and their catalytic mechanism and potential application. Nat Commun 12(1):1263

    Article  CAS  Google Scholar 

  5. Pai M, Crowther MA (2012) Neutralization of heparin activity. Handb Exp Pharmacol 207:265–277

    Article  CAS  Google Scholar 

  6. McCarthy CP, Vaduganathan M, Solomon E et al (2020) Running thin: implications of a heparin shortage. Lancet 395(10223):534–536

    Article  Google Scholar 

  7. Tang N, Bai H, Chen X et al (2020) Anticoagulant treatment is associated with decreased mortality in severe coronavirus disease 2019 patients with coagulopathy. J Thromb Haemost 18(5):1094–1099

    Article  CAS  Google Scholar 

  8. Gama CI, Hsieh-Wilson LC (2005) Chemical approaches to deciphering the glycosaminoglycan code. Curr Opin Chem Biol 9(6):609–619

    Article  CAS  Google Scholar 

  9. Li JP, Kusche-Gullberg M (2016) Heparan sulfate: biosynthesis, structure, and function. Int Rev Cell Mol Biol 325:215–273

    Article  CAS  Google Scholar 

  10. Casu B, Naggi A, Torri G (2015) Re-visiting the structure of heparin. Carbohydr Res 403:60–68

    Article  CAS  Google Scholar 

  11. Miller RL, Guimond SE, Schwörer R et al (2020) Shotgun ion mobility mass spectrometry sequencing of heparan sulfate saccharides. Nat Commun 11(1):1481

    Article  CAS  Google Scholar 

  12. Kozlowski AM, Yates EA, Roubroeks JP et al (2021) Hydrolytic degradation of heparin in acidic environments: nuclear magnetic resonance reveals details of selective desulfation. ACS Appl Mater Interfaces 13(4):5551–5563

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. School of Life Science and Technology, Weifang Medical University, Weifang, China

    Qingdong Zhang & Danrong Lu

  2. National Glycoengineering Research Center and Shandong Key Laboratory of Carbohydrate Chemistry and Glycobiology, Shandong University, Qingdao, China

    Fuchuan Li

Authors
  1. Qingdong Zhang
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  2. Danrong Lu
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  3. Fuchuan Li
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Corresponding author

Correspondence to Fuchuan Li .

Editor information

Editors and Affiliations

  1. Biochemistry, Biochemical Analysis and Matrix Pathobiology Research Group, Laboratory of Biochemistry, Department of Chemistry, University of Patras, Patras, Greece

    Nikos K. Karamanos

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Zhang, Q., Lu, D., Li, F. (2023). Enzymatic Sequencing of Heparin Oligosaccharides Using Exolyase. In: Karamanos, N.K. (eds) Proteoglycans. Methods in Molecular Biology, vol 2619. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2946-8_18

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  • DOI: https://doi.org/10.1007/978-1-0716-2946-8_18

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2945-1

  • Online ISBN: 978-1-0716-2946-8

  • eBook Packages: Springer Protocols

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