Skip to main content
SpringerLink
Log in

Isolation and Characterization of Heparan Sulfate Containing Amyloid Precursor Protein Degradation Products

  • Protocol
  • First Online:
Glycosaminoglycans

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2303))

Abstract

Numerous studies indicate that heparan sulfate proteoglycans (HSPGs) participate in a network of complex molecular events involving amyloid precursor protein (APP) processing and formation, oligomerization, intracellular targeting, clearance, and propagation of amyloid β in Alzheimer’s disease (AD). A mutual functional interplay between recycling glypican-1 and APP processing has been demonstrated where the HS released from glypican-1 by a Cu/NO-ascorbate-dependent reaction forms a conjugate with APP degradation products and undergoes an endosome-nucleus-autophagosome co-trafficking. HS has been shown to display contradictory and dual effects in AD involving both prevention and promotion of amyloid β formation. It is therefore important to identify the source, detailed structural features as well as factors that favor formation of the neuroprotective forms of HS. Here, a method for isolation and identification of HS-containing APP degradation products has been described. The method is based on isolation of radiolabeled HS followed by identification of accompanying APP degradation products by SDS-PAGE and Western blotting.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 329.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Jen YH, Musacchio M, Lander AD (2009) Glypican-1 controls brain size through regulation of fibroblast growth factor signaling in early neurogenesis. Neural Dev 4:33

    Article  Google Scholar 

  2. Ohmi K, Zhao HZ, Neufeld EF (2011) Defects in the medial entorhinal cortex and dentate gyrus in the mouse model of Sanfilippo syndrome type B. PLoS One 6:e27461

    Article  CAS  Google Scholar 

  3. Liebsch F, Kulic L, Teunissen C, Shobo A, Ulku I, Engelschalt V, Hancock MA, van der Flier WM, Kunach P, Rosa-Neto P, Scheltens P, Poirier J, Saftig P, Bateman RJ, Breitner J, Hock C, Multhaup G (2019) Aβ34 is a BACE1-derived degradation intermediate associated with amyloid clearance and Alzheimer's disease progression. Nat Commun 10:2240

    Article  Google Scholar 

  4. Wang J, Gu BJ, Masters CL, Wang YJ (2017) A systemic view of Alzheimer disease - insights from amyloid-β metabolism beyond the brain. Nat Rev Neurol 13:612–623

    Article  CAS  Google Scholar 

  5. Williamson TG, Mok SS, Henry A, Cappai R, Lander AD, Nurcombe V, Beyreuther K, Masters CL, Small DH (1996) Secreted glypican binds to the amyloid precursor protein of Alzheimer’s disease (APP) and inhibits APP-induced neurite outgrowth. J Biol Chem 271:31215–31221

    Article  CAS  Google Scholar 

  6. Xue Y, Lee S, Ha Y (2011) Crystal structure of amyloid precursor-like protein 1 and heparin complex suggests a dual role of heparin in E2 dimerization. Proc Natl Acad Sci U S A 108:16229–16234

    Article  CAS  Google Scholar 

  7. Dahms SO, Mayer MC, Roeser D, Multhaup G, Than ME (2015) Interaction of the amyloid precursor protein-like protein 1 (APLP1) E2 domain with heparan sulfate involves two distinct binding modes. Acta Crystallogr D Biol Crystallogr 71(Pt 3):494–504

    Article  CAS  Google Scholar 

  8. Reinhard C, Borgers M, David G, De Strooper B (2013) Soluble amyloid-β precursor protein binds its cell surface receptor in a cooperative fashion with glypican and syndecan proteoglycans. J Cell Sci 126:4856–4861

    CAS  PubMed  Google Scholar 

  9. Stopschinski BE, Holmes BB, Miller GM, Manon VA, Vaquer-Alicea J, Prueitt WL, Hsieh-Wilson LC, Diamond MI (2018) Specific glycosaminoglycan chain length and sulfation patterns are required for cell uptake of tau versus α-synuclein and β-amyloid aggregates. J Biol Chem 293:10826–10840

    Article  CAS  Google Scholar 

  10. Löfgren K, Cheng F, Fransson L-Å, Bedecs K, Mani K (2008) Involvement of glypican-1 autoprocessing in scrapie infection. Eur J Neurosci 28:964–972

    Article  Google Scholar 

  11. Cheng F, Cappai R, Fransson L-Å, Mani K (2014) APP/APLP2 expression is required to initiate endosome-nucleus-autophagosome trafficking of glypican-1-derived heparan sulfate. J Biol Chem 289:20871–20878

    Article  CAS  Google Scholar 

  12. Mani K, Cheng F, Havsmark B, David S, Fransson L-Å (2004) Involvement of glycosylphosphatidylinositol-linked ceruloplasmin in the copper/zinc-nitric oxide-dependent degradation of glypican-1 heparan sulfate in rat C6 glioma cells. J Biol Chem 279:12918–12923

    Article  CAS  Google Scholar 

  13. Cappai R, Cheng F, Ciccotosto GD, Needham BE, Masters CL, Multhaup G, Fransson L-Å, Mani K (2005) The amyloid precursor protein (APP) of Alzheimer disease and its paralog, APLP2, modulate the Cu/Zn-nitric oxide-catalyzed degradation of glypican-1 heparan sulfate in vivo. J Biol Chem 280:13913–13920

    Article  CAS  Google Scholar 

  14. Cheng F, Fransson L-Å, Mani K (2017) Cytochrome b561, copper, β-cleaved amyloid precursor protein and niemann-pick C1 protein are involved in ascorbate-induced release and membrane penetration of heparan sulfate from endosomal S-nitrosylated glypican-1. Exp Cell Res 360:171–179

    Article  CAS  Google Scholar 

  15. Mani K, Cheng F, Havsmark B, Jönsson M, Belting M, Fransson LA (2003) Prion, amyloid beta-derived Cu(II) ions, or free Zn(II) ions support S-nitroso-dependent autocleavage of glypican-1 heparan sulfate. J Biol 278:38956–38965

    CAS  Google Scholar 

  16. Mani K, Cheng F, Fransson L-Å (2007) Heparan sulfate degradation products can associate with oxidized proteins and proteasomes. J Biol Chem 282:21934–21944

    Article  CAS  Google Scholar 

  17. Cheng F, Belting M, Fransson L-Å, Mani K (2017) Nucleolin is a nuclear target of heparan sulfate derived from glypican-1. Exp Cell Res 354:31–39

    Article  CAS  Google Scholar 

  18. Cheng F, Fransson L-Å, Mani K (2015) Rapid nuclear transit and impaired degradation of amyloid beta and glypican-1-derived heparan sulfate in Tg2576 mouse fibroblasts. Glycobiology 25(5):548–556

    Article  CAS  Google Scholar 

  19. Cheng F, Fransson LÅ, Mani K (2019) The cyanobacterial neurotoxin β-N-methylamino-l-alanine prevents addition of heparan sulfate to glypican-1 and increases processing of amyloid precursor protein in dividing neuronal cells. Exp Cell Res 379:172–181

    Article  CAS  Google Scholar 

  20. van Horssen J, Kleinnijenhuis J, Maass CN, Rensink AA, Otte-Holler I, David G, van den Heuvel LP, Wesseling P, de Waal RM, Verbeek MM (2002) Accumulation of heparan sulfate proteoglycans in cerebellar senile plaques. Neurobiol Aging 23:537–545

    Article  Google Scholar 

  21. Timmer NM, van Horssen J, Otte-Holler I, Wilhelmus MM, David G, van Beers J, de Waal RM, Verbeek MM (2009) Amyloid beta induces cellular relocalization and production of agrin and glypican-1. Brain Res 1260:38–46

    Article  CAS  Google Scholar 

  22. Cheng F, Cappai R, Ciccotosto GD, Svensson G, Multhaup G, Fransson L-Å, Mani K (2011) Suppression of amyloid beta A11 antibody immunoreactivity by vitamin C: possible role of heparan sulfate oligosaccharides derived from glypican-1 by ascorbate-induced, nitric oxide (NO)-catalyzed degradation. J Biol Chem 286:27559–27572

    Article  CAS  Google Scholar 

  23. Liu CC, Zhao N, Yamaguchi Y, Cirrito JR, Kanekiyo T, Holtzman DM, Bu G (2016) Neuronal heparan sulfates promote amyloid pathology by modulating brain amyloid-β clearance and aggregation in Alzheimer's disease. Sci Transl Med 8:332ra44

    PubMed  PubMed Central  Google Scholar 

  24. Schwörer R, Zubkova OV, Turnbull JE, Tyler PC (2013) Synthesis of a targeted library of heparan sulfate hexa- to dodecasaccharides as inhibitors of β-secretase: potential therapeutics for Alzheimer’s disease. Chemistry 19:6817–6823

    Article  Google Scholar 

  25. Li J, Li J, Sun T, Cai C, Shao M, Yu G (2019) Concise chemoenzymatic synthesis of heparan sulfate analogues as potent BACE-1 inhibitors. Carbohydr Polym 217:232–239

    Article  CAS  Google Scholar 

  26. García B, Martín C, García-Suárez O, Muñiz-Alonso B, Ordiales H, Fernández-Menéndez S, Santos-Juanes J, Lorente-Gea L, Castañón S, Vicente-Etxenausia I, Piña Batista KM, Ruiz-Díaz I, Caballero-Martínez MC, Merayo-Lloves J, Guerra-Merino I, Quirós LM, Fernández-Vega I (2017) Upregulated expression of Heparanase and Heparanase 2 in the brains of Alzheimer’s disease. J Alzheimers Dis 58:185–192

    Article  Google Scholar 

  27. Jendresen CB, Cui H, Zhang X, Vlodavsky I, Nilsson LN, Li JP (2015) Overexpression of heparanase lowers the amyloid burden in amyloid-β precursor protein transgenic mice. J Biol Chem 290:5053–5064

    Article  CAS  Google Scholar 

  28. Small DH, Williamson T, Reed G, Clarris H, Beyreuther K, Masters CL, Nurcombe V (1996) The role of heparan sulfate proteoglycans in the pathogenesis of Alzheimer's disease. Ann N Y Acad Sci 777:316–321

    Article  CAS  Google Scholar 

  29. Reilly CE (2000) Crucial role of heparan sulfate proteoglycan (agrin) in beta-amyloid formation in Alzheimer’s disease. J Neurol 247:663–664

    Article  CAS  Google Scholar 

  30. Scholefield Z, Yates EA, Wayne G, Amour A, McDowell W, Turnbull JE (2003) Heparan sulfate regulates amyloid precursor protein processing by BACE1, the Alzheimer’s beta-secretase. J Cell Biol 163:97–107

    Article  CAS  Google Scholar 

  31. Beckman M, Holsinger RM, Small DH (2006) Heparin activates beta-secretase (BACE1) of Alzheimer’s disease and increases autocatalysis of the enzyme. Biochemistry 45:6703–6714

    Article  CAS  Google Scholar 

  32. Small DH, Klaver DW, Beckman M (2008) Regulation of proBACE1 by glycosaminoglycans. Neurodegener Dis 5:206–208

    Article  CAS  Google Scholar 

  33. Kanekiyo T, Zhang J, Liu Q, Liu CC, Zhang L, Bu G (2011) Heparan sulphate proteoglycan and the low-density lipoprotein receptor-related protein 1 constitute major pathways for neuronal amyloid-beta uptake. J Neurosci 31:1644–1651

    Article  CAS  Google Scholar 

  34. Wesén E, Gallud A, Paul A, Lindberg DJ, Malmberg P, Esbjörner EK (2018) Cell surface proteoglycan-mediated uptake and accumulation of the Alzheimer's disease peptide Aβ (1-42). Biochim Biophys Acta Biomembr 1860:2204–2214

    Article  Google Scholar 

  35. Cheng F, Ruscher K, Fransson LÅ, Mani K (2013) Non-toxic amyloid beta formed in the presence of glypican-1 or its deaminatively generated heparan sulfate degradation products. Glycobiology 23:1510–1519

    Article  CAS  Google Scholar 

  36. O'Callaghan P, Sandwall E, Li JP, Yu H, Ravid R, Guan ZZ, van Kuppevelt TH, Nilsson LN, Ingelsson M, Hyman BT, Kalimo H, Lindahl U, Lannfelt L, Zhang X (2008) Heparan sulfate accumulation with Abeta deposits in Alzheimer’s disease and Tg2576 mice is contributed by glial cells. Brain Pathol 18:548–561

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Bruinsma IB, te Riet L, Gevers T, ten Dam GB, van Kuppevelt TH, David G, Küsters B, de Waal RM, Verbeek MM (2010) Sulfation of heparan sulfate associated with amyloid-beta plaques in patients with Alzheimer’s disease. Acta Neuropathol 119:211–220

    Article  CAS  Google Scholar 

  38. Sandwall E, O'Callaghan P, Zhang X, Lindahl U, Lannfelt L, Li JP (2010) Heparan sulfate mediates amyloid-beta internalization and cytotoxicity. Glycobiology 20:533–5341

    Article  CAS  Google Scholar 

  39. Castillo GM, Lukito W, Wight TN, Snow AD (1999) The sulfate moieties of glycosaminoglycans are critical for the enhancement of beta-amyloid protein fibril formation. J Neurochem 72:1681–1687

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Experimental Medical Science, Division of Neuroscience, Glycobiology Group, Lund University, Lund, Sweden

    Katrin Mani

Authors
  1. Katrin Mani
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Katrin Mani .

Editor information

Editors and Affiliations

  1. Departments of Biology, Bioengineering, and Medicinal Chemistry, University of Utah, Salt Lake City, UT, USA

    Kuberan Balagurunathan

  2. Department of Genetics, Cell Biology and Development, University of Minnesota, Minneapolis, MN, USA

    Hiroshi Nakato

  3. Department of Medicinal Chemistry and Institute for Structural Biology, Drug Discovery and Development, Virginia Commonwealth University, Richmond, VA, USA

    Umesh Desai

  4. Department of Neurobiology & Department of Nutrition & Integrative Physiology, University of Utah, Salt Lake City, UT, USA

    Yukio Saijoh

Rights and permissions

Reprints and permissions

Copyright information

© 2022 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Mani, K. (2022). Isolation and Characterization of Heparan Sulfate Containing Amyloid Precursor Protein Degradation Products. In: Balagurunathan, K., Nakato, H., Desai, U., Saijoh, Y. (eds) Glycosaminoglycans. Methods in Molecular Biology, vol 2303. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1398-6_22

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-1398-6_22

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1397-9

  • Online ISBN: 978-1-0716-1398-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation