Skip to main content

ZG16p, an Animal Homologue of Plant β-Prism Fold Lectins: Purification Methods of Natural and Recombinant ZG16p and Inhibition Assay of Cancer Cell Growth Using ZG16p

  • Protocol
  • First Online:
Lectin Purification and Analysis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2132))

Abstract

ZG16p is a soluble 16-kDa protein abundantly expressed in the pancreas and gut, and has a β-prism fold structure similar to that of mannose-binding Jacalin-related lectins (mJRLs) such as BanLec, Heltuba, and Artocarpin. ZG16p binds to mannose via the well-conserved GXXXD loop among mJRLs and sulfated glycosaminoglycans (e.g., heparin and heparan sulfate) via the basic patch of molecular surface. In addition to the above binding activities, ZG16p has inhibitory activity against proliferation of colon cancer cells. This manuscript describes purification of rat pancreatic ZG16p and recombinant ZG16p expressed in Escherichia coli expression system, and cell growth inhibition assay using ZG16p as an inhibitor.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 189.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 239.00
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. Cronshagen U, Voland P, Kern HF (1994) cDNA cloning and characterization of a novel 16 kDa protein located in zymogen granules of rat pancreas and goblet cells of the gut. Eur J Cell Biol 65:366–377

    CAS  PubMed  Google Scholar 

  2. Kumazawa-Inoue K, Mimura T, Hosokawa-Tamiya S et al (2012) ZG16p, an animal homologue of β-prism fold plant lectins, interacts with heparan sulfate proteoglycans in pancreatic zymogen granules. Glycobiology 22:258–266

    Article  CAS  Google Scholar 

  3. Kleene R, Dartsch H, Kern HF (1999) The secretory lectin ZG16p mediates sorting of enzyme proteins to the zymogen granule membrane in pancreatic acinar cells. Eur J Cell Biol 78:79–90

    Article  CAS  Google Scholar 

  4. Schmidt K, Dartsch H, Linder D et al (2000) A submembranous matrix of proteoglycans on zymogen granule membranes is involved in granule formation in rat pancreatic acinar cells. J Cell Sci 113:2233–2242

    CAS  PubMed  Google Scholar 

  5. Tateno H, Yabe R, Sato T et al (2012) Human ZG16p recognizes pathogenic fungi through non-self polyvalent mannose in the digestive system. Glycobiology 22:210–220

    Article  CAS  Google Scholar 

  6. Kanagawa M, Satoh T, Ikeda A et al (2011) Crystal structures of human secretory proteins ZG16p and ZG16b reveal a Jacalin-related β-prism fold. Biochem Biophys Res Commun 404:201–205

    Article  CAS  Google Scholar 

  7. Meagher JL (2005) Crystal structure of banana lectin reveals a novel second sugar binding site. Glycobiology 15:1033–1042

    Article  CAS  Google Scholar 

  8. Kanagawa M, Liu Y, Hanashima S et al (2014) Structural basis for multiple sugar recognition of Jacalin-related human ZG16p lectin. J Biol Chem 289:16954–16965

    Article  CAS  Google Scholar 

  9. Singh R (2006) Peanut lectin stimulates proliferation of colon cancer cells by interaction with glycosylated CD44v6 isoforms and consequential activation of c-met and MAPK: functional implications for disease-associated glycosylation changes. Glycobiology 16:594–601

    Article  CAS  Google Scholar 

  10. Deepa M, Sureshkumar T, Satheeshkumar PK et al (2012) Purified mulberry leaf lectin (MLL) induces apoptosis and cell cycle arrest in human breast cancer and colon cancer cells. Chem Biol Interact 200:38–44

    Article  CAS  Google Scholar 

  11. Mori Y, Akita K, Yashiro M et al (2015) Binding of galectin-3, a β-galactoside-binding lectin, to MUC1 protein enhances phosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2) and Akt, promoting tumor cell malignancy. J Biol Chem 290:26125–26140

    Article  CAS  Google Scholar 

  12. Satelli A, Rao PS, Thirumala S et al (2011) Galectin-4 functions as a tumor suppressor of human colorectal cancer. Int J Cancer 129:799–809

    Article  CAS  Google Scholar 

  13. Mito A, Nakano Y, Saitoh T et al (2018) Lectin ZG16p inhibits proliferation of human colorectal cancer cells via its carbohydrate-binding sites. Glycobiology 28:21–31

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kyoko Kojima-Aikawa .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Mito, A., Kumazawa-Inoue, K., Kojima-Aikawa, K. (2020). ZG16p, an Animal Homologue of Plant β-Prism Fold Lectins: Purification Methods of Natural and Recombinant ZG16p and Inhibition Assay of Cancer Cell Growth Using ZG16p. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_33

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_33

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0429-8

  • Online ISBN: 978-1-0716-0430-4

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics