Identification of a novel cytotoxic protein, Cry45Aa, from Bacillus thuringiensis A1470 and its selective cytotoxic activity against various mammalian cell lines

Shiro Okumura, Hiroyuki Saitoh, Tomoyuki Ishikawa, Naoya Wasano, Satoko Yamashita, Ken Ichi Kusumoto, Tetsuyuki Akao, Eiichi Mizuki, Michio Ohba, Kuniyo Inouye

Research output: Contribution to journalArticlepeer-review

26 Citations (Scopus)

Abstract

Parasporal inclusion proteins produced by Bacillus thuringiensis strain A1470 exhibit strong cytotoxicity against human leukemic T cells when activated by protease treatment. One of the cytotoxic proteins was separated by anion exchange chromatography and gel filtration chromatography and designated Cry45Aa. Its gene was then expressed in recombinant Escherichia coli, in which the Cry45Aa precursor was accumulated in an inclusion body. It was solubilized in sodium carbonate buffer and processed with proteinase K, and cytotoxic activities of the protein against various mammalian cell lines were evaluated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide assay. The protein exhibited high cytotoxic activity against CACO-2, Sawano, MOLT-4, TCS, and HL60 cells and moderate activity against U-937 DE-4, PC12, and HepG2 cells. On the other hand, the EC50 values against Jurkat, K562, HeLa, A549, Vero, COS-7, NIH3T3, CHO, and four normal tissue cells (human primary hepatocyte cells, UtSMC, MRC-5, and normal T cells) were >2 μg/mL.

Original languageEnglish
Pages (from-to)6313-6318
Number of pages6
JournalJournal of Agricultural and Food Chemistry
Volume53
Issue number16
DOIs
Publication statusPublished - Aug 10 2005
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

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