Characterization of Functional Domains of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata

Yoshiaki Kouzuma, Yota Suzuki, Masahiro Nakano, Kayo Matsuyama, Sumiki Tojo, Makoto Kimura, Takayuki Yamasaki, Haruhiko Aoyagi, Tomomitsu Hatakeyama

Research output: Contribution to journalArticle

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Abstract

CEL-III is a Ca2+-dependent, galactose/N-acetylgalactosamine (GalNAc)-specific lectin isolated from the marine invertebrate Cucumaria echinata. This lectin exhibits strong hemolytic activity and cytotoxicity through pore formation in target cell membranes. The amino acid sequence of CEL-III revealed the N-terminal two-thirds to have homology to the B-chains of ricin and abrin, which are galactose-specific plant toxic lectins; the C-terminal one-third shows no homology to any known proteins. To examine the carbohydrate-binding ability of the N-terminal region of CEL-III, the protein comprising Pyr1-Phe283 was expressed in Escherichia coli cells. The expressed protein showed both the ability to bind to a GalNAc-immobilized column as well as hemagglutinating activity for rabbit erythrocytes, confirming that the N-terminal region has binding activity for specific carbohydrates. Since the C-terminal region could not be expressed in E. coli cells, a fragment containing this region was produced by limited proteolysis of the native protein by trypsin. The resulting C-terminal 15 kDa fragment of CEL-III exhibited a tendency to self-associate, forming an oligomer. When mixed with erythrocytes, the oligomer of the C-terminal fragment caused hemagglutination, probably due to hydrophobic interaction with cell membranes, while the monomeric fragment did not. Chymotryptic digestion of the preformed CEL-III oligomer induced upon lactose binding also yielded an oligomer of the C-terminal fragment comprising six molecules of the 16 kDa fragment. These results suggest that after binding to cell surface carbohydrate chains, CEL-III oligomerizes through C-terminal domains, leading to the formation of ion-permeable pores by hydrophobic interaction with the cell membrane.

Original languageEnglish
Pages (from-to)395-402
Number of pages8
JournalJournal of biochemistry
Volume134
Issue number3
DOIs
Publication statusPublished - Sep 1 2003

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Cucumaria
Invertebrates
Oligomers
Cell membranes
Galactose
Lectins
Acetylgalactosamine
Carbohydrates
Cell Membrane
Hydrophobic and Hydrophilic Interactions
Escherichia coli
Abrin
Proteins
Erythrocytes
Plant Lectins
Proteolysis
Ricin
Poisons
Hemagglutination
Lactose

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Kouzuma, Y., Suzuki, Y., Nakano, M., Matsuyama, K., Tojo, S., Kimura, M., ... Hatakeyama, T. (2003). Characterization of Functional Domains of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata. Journal of biochemistry, 134(3), 395-402. https://doi.org/10.1093/jb/mvg157

Characterization of Functional Domains of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata. / Kouzuma, Yoshiaki; Suzuki, Yota; Nakano, Masahiro; Matsuyama, Kayo; Tojo, Sumiki; Kimura, Makoto; Yamasaki, Takayuki; Aoyagi, Haruhiko; Hatakeyama, Tomomitsu.

In: Journal of biochemistry, Vol. 134, No. 3, 01.09.2003, p. 395-402.

Research output: Contribution to journalArticle

Kouzuma, Y, Suzuki, Y, Nakano, M, Matsuyama, K, Tojo, S, Kimura, M, Yamasaki, T, Aoyagi, H & Hatakeyama, T 2003, 'Characterization of Functional Domains of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata', Journal of biochemistry, vol. 134, no. 3, pp. 395-402. https://doi.org/10.1093/jb/mvg157
Kouzuma, Yoshiaki ; Suzuki, Yota ; Nakano, Masahiro ; Matsuyama, Kayo ; Tojo, Sumiki ; Kimura, Makoto ; Yamasaki, Takayuki ; Aoyagi, Haruhiko ; Hatakeyama, Tomomitsu. / Characterization of Functional Domains of the Hemolytic Lectin CEL-III from the Marine Invertebrate Cucumaria echinata. In: Journal of biochemistry. 2003 ; Vol. 134, No. 3. pp. 395-402.
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