Skip to main content

Galactose-Specific, Hemolytic Lectin CEL-III from Cucumaria echinata

  • Protocol
  • First Online:
Book cover Lectin Purification and Analysis

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

Abstract

CEL-III is a Ca2+-dependent and galactose-specific lectin purified from the sea cucumber, Cucumaria echinata; it exhibits hemolytic and hemagglutinating activities. CEL-III consists of the following three distinct domains: two N-terminal carbohydrate-binding domains (1 and 2), which adopt β-trefoil folds such as the B-chain of ricin and are members of the (QXW)3 motif family, and domain 3, an oligomerization domain. After binding to the cell surface carbohydrate chains through domains 1 and 2, domain 3 self-associates to form transmembrane pores composed of CEL-III heptamers, leading to cell lysis or death. In this chapter, the purification and carbohydrate-coated microplate binding assay of CEL-III are described.

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. Hatakeyama T, Kohzaki H, Nagatomo H, Yamasaki N (1994) Purification and characterization of four Ca2+-dependent lectins from the marine invertebrate, Cucumaria echinata. J Biochem 116:209–214

    Article  CAS  Google Scholar 

  2. Hatakeyama T, Ohuchi K, Kuroki M, Yamasaki N (1995) Amino acid sequence of a C-type lectin CEL-IV from the marine invertebrate Cucumaria echinata. Biosci Biotechnol Biochem 59:1314–1317

    Article  CAS  Google Scholar 

  3. Hatakeyama T, Matsuo N, Shiba K et al (2002) Amino acid sequence and carbohydrate-binding analysis of the N-acetyl-D-galactosamine-specific C-type lectin, CEL-I, from the Holothuroidea, Cucumaria echinata. Biosci Biotechnol Biochem 66:157–163

    Article  CAS  Google Scholar 

  4. Hatakeyama T, Nagatomo H, Yamasaki N (1995) Interaction of the hemolytic lectin CEL-III from the marine invertebrate Cucumaria echinata with the erythrocyte membrane. J Biol Chem 270:3560–3564

    Article  CAS  Google Scholar 

  5. Oda T, Tsuru M, Hatakeyama T et al (1997) Temperature- and pH-dependent cytotoxic effect of the hemolytic lectin CEL-III from the marine invertebrate Cucumaria echinata on various cell lines. J Biochem 121:560–567

    Article  CAS  Google Scholar 

  6. Hatakeyama T, Furukawa M, Nagatomo H et al (1996) Oligomerization of the hemolytic lectin CEL-III from the marine invertebrate Cucumaria echinata induced by the binding of carbohydrate ligands. J Biol Chem 271:16915–16920

    Article  CAS  Google Scholar 

  7. Kuwahara H, Yamasaki T, Hatakeyama T et al (2002) Oligomerization process of the hemolytic lectin CEL-III purified from a sea cucumber, Cucumaria echinata. J Biochem 131:751–756

    Article  CAS  Google Scholar 

  8. Uchida T, Yamasaki T, Eto S et al (2004) Crystal structure of the hemolytic lectin CEL-III isolated from the marine invertebrate Cucumaria echinata: implications of domain structure for its membrane pore-formation mechanism. J Biol Chem 279:37133–37141

    Article  CAS  Google Scholar 

  9. Hatakeyama T, Unno H, Kouzuma Y et al (2007) C-type lectin-like carbohydrate recognition of the hemolytic lectin CEL-III containing ricin-type β-trefoil folds. J Biol Chem 282:37826–37835

    Article  CAS  Google Scholar 

  10. Unno H, Hisamatsu K, Nagao T et al (2013) Crystallization and preliminary crystallographic study of oligomers of the haemolytic lectin CEL-III from the sea cucumber Cucumaria echinata. Acta Crystallogr Sect F Struct Biol Cryst Commun 69:416–420

    Article  CAS  Google Scholar 

  11. Hatakeyama T, Sato T, Taira E et al (1999) Characterization of the interaction of hemolytic lectin CEL-III from the marine invertebrate, Cucumaria echinata, with artificial lipid membranes: involvement of neutral sphingoglycolipids in the pore-forming process. J Biochem 125:277–284

    Article  CAS  Google Scholar 

  12. Hatakeyama T, Miyamoto Y, Nagatomo H et al (1997) Carbohydrate-binding properties of the hemolytic lectin CEL-III from the holothuroidea Cucumaria echinata as analyzed using carbohydrate-coated microplate. J Biochem 121:63–67

    Article  CAS  Google Scholar 

  13. Teichberg VI, Aberdam D, Erez U, Pinelli E (1988) Affinity-repulsion chromatography. Principle and application to lectins. J Biol Chem 263:14086–14092

    CAS  PubMed  Google Scholar 

  14. Ciesiolka T, Gabius HJ (1988) An 8- to 10-fold enhancement in sensitivity for quantitation of proteins by modified application of colloidal gold. Anal Biochem 168:280–283

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Tomomitsu Hatakeyama .

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

Hatakeyama, T. (2020). Galactose-Specific, Hemolytic Lectin CEL-III from Cucumaria echinata. 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_16

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0430-4_16

  • 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