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Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain

Parasporin is the cytocidal protein present in the parasporal inclusion of the non-insecticidal Bacillus thuringiensis strains, which has no hemolytic activity but has cytocidal activities, preferentially killing cancer cells. In this study, we characterized a cytocidal protein that belongs to this...

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Autores principales: Ekino, Keisuke, Okumura, Shiro, Ishikawa, Tomoyuki, Kitada, Sakae, Saitoh, Hiroyuki, Akao, Tetsuyuki, Oka, Takuji, Nomura, Yoshiyuki, Ohba, Michio, Shin, Takashi, Mizuki, Eiichi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4073135/
https://www.ncbi.nlm.nih.gov/pubmed/24945755
http://dx.doi.org/10.3390/toxins6061882
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author Ekino, Keisuke
Okumura, Shiro
Ishikawa, Tomoyuki
Kitada, Sakae
Saitoh, Hiroyuki
Akao, Tetsuyuki
Oka, Takuji
Nomura, Yoshiyuki
Ohba, Michio
Shin, Takashi
Mizuki, Eiichi
author_facet Ekino, Keisuke
Okumura, Shiro
Ishikawa, Tomoyuki
Kitada, Sakae
Saitoh, Hiroyuki
Akao, Tetsuyuki
Oka, Takuji
Nomura, Yoshiyuki
Ohba, Michio
Shin, Takashi
Mizuki, Eiichi
author_sort Ekino, Keisuke
collection PubMed
description Parasporin is the cytocidal protein present in the parasporal inclusion of the non-insecticidal Bacillus thuringiensis strains, which has no hemolytic activity but has cytocidal activities, preferentially killing cancer cells. In this study, we characterized a cytocidal protein that belongs to this category, which was designated parasporin-5 (PS5). PS5 was purified from B. thuringiensis serovar tohokuensis strain A1100 based on its cytocidal activity against human leukemic T cells (MOLT-4). The 50% effective concentration (EC(50)) of PS5 to MOLT-4 cells was approximately 0.075 μg/mL. PS5 was expressed as a 33.8-kDa inactive precursor protein and exhibited cytocidal activity only when degraded by protease at the C-terminal into smaller molecules of 29.8 kDa. Although PS5 showed no significant homology with other known parasporins, a Position Specific Iterative-Basic Local Alignment Search Tool (PSI-BLAST) search revealed that the protein showed slight homology to, not only some B. thuringiensis Cry toxins, but also to aerolysin-type β-pore-forming toxins (β-PFTs). The recombinant PS5 protein could be obtained as an active protein only when it was expressed in a precursor followed by processing with proteinase K. The cytotoxic activities of the protein against various mammalian cell lines were evaluated. PS5 showed strong cytocidal activity to seven of 18 mammalian cell lines tested, and low to no cytotoxicity to the others.
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spelling pubmed-40731352014-06-27 Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain Ekino, Keisuke Okumura, Shiro Ishikawa, Tomoyuki Kitada, Sakae Saitoh, Hiroyuki Akao, Tetsuyuki Oka, Takuji Nomura, Yoshiyuki Ohba, Michio Shin, Takashi Mizuki, Eiichi Toxins (Basel) Article Parasporin is the cytocidal protein present in the parasporal inclusion of the non-insecticidal Bacillus thuringiensis strains, which has no hemolytic activity but has cytocidal activities, preferentially killing cancer cells. In this study, we characterized a cytocidal protein that belongs to this category, which was designated parasporin-5 (PS5). PS5 was purified from B. thuringiensis serovar tohokuensis strain A1100 based on its cytocidal activity against human leukemic T cells (MOLT-4). The 50% effective concentration (EC(50)) of PS5 to MOLT-4 cells was approximately 0.075 μg/mL. PS5 was expressed as a 33.8-kDa inactive precursor protein and exhibited cytocidal activity only when degraded by protease at the C-terminal into smaller molecules of 29.8 kDa. Although PS5 showed no significant homology with other known parasporins, a Position Specific Iterative-Basic Local Alignment Search Tool (PSI-BLAST) search revealed that the protein showed slight homology to, not only some B. thuringiensis Cry toxins, but also to aerolysin-type β-pore-forming toxins (β-PFTs). The recombinant PS5 protein could be obtained as an active protein only when it was expressed in a precursor followed by processing with proteinase K. The cytotoxic activities of the protein against various mammalian cell lines were evaluated. PS5 showed strong cytocidal activity to seven of 18 mammalian cell lines tested, and low to no cytotoxicity to the others. MDPI 2014-06-18 /pmc/articles/PMC4073135/ /pubmed/24945755 http://dx.doi.org/10.3390/toxins6061882 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Ekino, Keisuke
Okumura, Shiro
Ishikawa, Tomoyuki
Kitada, Sakae
Saitoh, Hiroyuki
Akao, Tetsuyuki
Oka, Takuji
Nomura, Yoshiyuki
Ohba, Michio
Shin, Takashi
Mizuki, Eiichi
Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain
title Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain
title_full Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain
title_fullStr Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain
title_full_unstemmed Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain
title_short Cloning and Characterization of a Unique Cytotoxic Protein Parasporin-5 Produced by Bacillus thuringiensis A1100 Strain
title_sort cloning and characterization of a unique cytotoxic protein parasporin-5 produced by bacillus thuringiensis a1100 strain
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4073135/
https://www.ncbi.nlm.nih.gov/pubmed/24945755
http://dx.doi.org/10.3390/toxins6061882
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