International Journal of Biological Sciences

Impact factor
3.873

ISSN 1449-2288

News feeds of IJBS published articles
My Manuscript
My Account

Journal of Biomedicinenew

Theranostics

International Journal of Medical Sciences

Journal of Cancer

Oncomedicine

Journal of Genomics

Journal of Bone and Joint Infection (JBJI)

Nanotheranostics

Journal of Genomics now in PubMed/PubMed Central. Submit manuscript...

PubMed Central Indexed in Journal Impact Factor

Int J Biol Sci 2011; 7(6):729-739. doi:10.7150/ijbs.7.729

Research Paper

Purification and Characterization of Gigantoxin-4, a New Actinoporin from the Sea Anemone Stichodactyla Gigantea

Bo Hu*, Wei Guo*, Liang-hua Wang*, Jian-guang Wang, Xiao-yu Liu, Bing-hua Jiao

Department of Biochemistry and Molecular Biology, Second Military Medical University, Shanghai 200433, China
* These authors made equal contributions to this work.

Abstract

A new Cytolysin, termed as Gigantoxin-4, was isolated from the sea anemone Stichodactyla gigantea and found to be highly homologous with Cytolysin-3 (HMg III) from Heteractis magnifica, RTX-A from Radianthus macrodactylus, and Sticholysin-1 (St I) and Sticholysin-2 (St II) from Stichodactyla helianthus (homology 82%, 86%, 82% and 86% respectively). Its 20 N-terminal residues were identified and the full-length cDNA sequence was obtained by reverse transcription-polymerase chain reaction (RT-PCR). Multiple sequence alignments with other Cytolysins of the actinoporin family clearly indicated that Gigantoxin-4 belongs to this protein family. SDS-PAGE electrophoresis showed that this new actinoporin had a molecular mass of about 19 kDa, and possessed a high hemolytic activity to human erythrocytes (HA50= 40 ng/ml), which was inhibited by pre-incubation with sphingomyelin (SM) or SM-cholesterol mixtures. Our in vivo experiments showed that Gigantoxin-4 had wide toxicity to the rat cardiovascular system and the respiratory system. A concentration of 30μg/kg Gigantoxin-4, i.v. produced a positive inotropic effect on the rat heart although final cardiovascular failure was inevitable, and 60μg/kg Gigantoxin-4 caused respiratory arrest rapidly resulting in rat death. HE staining indicated pathological changes in various organs and tissues after i.v. administration of Gigantoxin-4.

Keywords: Sea anemone, Stichodactyla gigantea, Actinoporin, Sphingomyelin, in vivo

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See http://ivyspring.com/terms for full terms and conditions.
How to cite this article:
Hu B, Guo W, Wang Lh, Wang Jg, Liu Xy, Jiao Bh. Purification and Characterization of Gigantoxin-4, a New Actinoporin from the Sea Anemone Stichodactyla Gigantea. Int J Biol Sci 2011; 7(6):729-739. doi:10.7150/ijbs.7.729. Available from http://www.ijbs.com/v07p0729.htm