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Characterization of a Novel Proteinous Toxin from Sea Anemone Actineria villosa

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Abstract

The sea anemone Actineria villosa expresses a lethal protein toxin. We isolated a novel 120-kDa protein, Avt120, from partially purified toxin and found it to possess extremely strong lethal activity. The 3,453-bp Avt120 gene translates to a 995-amino acid protein. The 50% lethal dose (LD50) of purified Avt120 in mice was 85.17 ng. Among several tested cell lines, Colo205 cells were most sensitive to Avt120: 50% of them were damaged by 38.4 ng/mL Avt120. Avt120 exerted ATP degradation activity (10 μmol ATP h−1 mg−1), which was strongly inhibited by ganglioside GM1 to decrease the cytotoxicity of Avt120.

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Abbreviations

ADP:

Adenosine diphosphate

ATP:

Adenosine triphosphate

DTT:

Dithiothreitol

FCS:

Fetal calf serum

HPLC:

High performance liquid chromatography

LD:

Lethal dose

MEM:

Minimal essential medium

MTT assay:

Methylthiazole tetrazolium assay

PBS:

Phosphate buffered saline

SDS-PAGE:

Sodium dodecyl sulfate–polyacrylamide gel electrophoresis

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Acknowledgments

We would like to thank Y. Araki for help with sea anemone collection and advice on evaluating the biological functions of the toxin. We also thank Dr. A. Takemura at Department of Chemistry, Biology and Marine Sciences, University of the Ryukyus, for supporting the amino acid sequence of Avt120.

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Authors and Affiliations

  1. Department of Virology, Institute of Tropical Medicine, Nagasaki University, 1-12-4 Sakamoto, Nagasaki City, Nagasaki, 852-8523, Japan

    Gen-ichiro Uechi

  2. Division of Tropical Parasitology, Unit of Social and Environmental Medicine, School of Medicine, University of the Ryukyus, Nishihara, Japan

    Hiromu Toma & Yoshiya Sato

  3. Division of Molecular Microbiology, Center of Molecular Biosciences, University of the Ryukyus, Nishihara, Japan

    Takeshi Arakawa

Authors
  1. Gen-ichiro Uechi
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  2. Hiromu Toma
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  3. Takeshi Arakawa
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  4. Yoshiya Sato
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Corresponding author

Correspondence to Gen-ichiro Uechi.

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Uechi, Gi., Toma, H., Arakawa, T. et al. Characterization of a Novel Proteinous Toxin from Sea Anemone Actineria villosa . Protein J 30, 422–428 (2011). https://doi.org/10.1007/s10930-011-9347-8

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  • DOI: https://doi.org/10.1007/s10930-011-9347-8

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