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J. Biol. Chem., Vol. 279, Issue 20, 21282-21286, May 14, 2004

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A Bacillus thuringiensis Crystal Protein with Selective Cytocidal Action to Human Cells^*

Akio Ito, Yasuyuki Sasaguri¶, Sakae Kitada, Yoshitomo Kusaka, Kyoko Kuwano, Kenjiro Masutomi, Eiichi Mizuki||, Tetsuyuki Akao||, and Michio Ohba**

From the Department of Chemistry, Faculty of Science, Kyushu University, Fukuoka 812-8581, the ^¶Department of Pathology and Cell Biology, University of Occupational and Environmental Health, Fukuoka 807-8555, the ^||Biotechnology and Food Research Institute, Fukuoka Industrial Technology Center, Fukuoka 839-0861, and the ^**Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University, Fukuoka 812-8581, Japan

Bacillus thuringiensis crystal proteins, well known to be toxic to certain insects but not pathogenic to mammals, are used as insecticidal proteins in agriculture and forest management. We here identified a crystal protein that is non-insecticidal and non-hemolytic but has strong cytocidal activity against various human cells with a markedly divergent target specificity, e.g. highly cytotoxic to HepG2 and Jurkat and less cytotoxic to the normal hepatocyte (HC) and HeLa. In slices of liver and colon cancer tissues, the toxin protein preferentially killed the cancer cells, leaving other cells unaffected. The cytocidal effect of the protein is non-apoptotic with swelling and fragmentation of the susceptible cells, although the apoptotic process does occur when the cell damage proceeded slowly. The amino acid sequence deduced from the nucleotide sequence of the cloned gene of the protein has little sequence homology with the insecticidal crystal proteins of B. thuringiensis. These observations raise the presence of a new group of the B. thuringiensis toxin and the possibility of new applications for the protein in the medical field.

Received for publication, February 20, 2004

The nucleotide sequence(s) reported in this paper has been submitted to the GenBank^TM/EBI Data Bank with accession number(s) AB099515.

^* The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. AB099515

To whom correspondence should be addressed. Fax: 81-92-861-5737; E-mail: a.itoscc{at}mbox.nc.kyushu-u.ac.jp.

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