Journal article
Transgenic Biosynthesis of Trypanothione Protects Escherichia coli from Radiation-Induced Toxicity
Radiation research, Vol.174(3), pp.290-296
09/2010
DOI: 10.1667/RR2235.1
PMCID: PMC2942016
PMID: 20726720
Abstract
Trypanothione is a unique diglutathionyl-spermidine conjugate found in abundance in trypanosomes but not in other eukaryotes. Because trypanothione is a naturally occurring polyamine thiol reminiscent of the synthetic drug amifostine, it may be a useful protector against radiation and oxidative stress. For these reasons we hypothesized that trypanothione might serve as a radioprotective agent when produced in bacteria. To accomplish this objective, the trypanothione synthetase and reductase genes from
T. cruzi
were introduced into
E. coli
and their expression was verified by qPCR and immunoblotting. Trypanothione synthesis in bacteria, detected by HPLC, resulted in decreased intracellular levels of reactive oxygen species as determined by H
2
DCFDA oxidation. Moreover,
E. coli
genomic DNA was protected from radiation-induced DNA damage by 4.6-fold in the presence of trypanothione compared to control bacteria. Concordantly, the transgenic
E. coli
expressing trypanothione were 4.3-fold more resistant to killing by
137
Cs γ radiation compared to
E. coli
devoid of trypanothione expression. Thus we have shown for the first time that
E. coli
can be genetically engineered to express the trypanothione biosynthetic pathway and produce trypanothione, which results in their radioresistance. These results warrant further research to explore the possibility of developing trypanothione as a novel radioprotective agent.
Details
- Title: Subtitle
- Transgenic Biosynthesis of Trypanothione Protects Escherichia coli from Radiation-Induced Toxicity
- Creators
- Matthew P Fitzgerald - The University of Iowa Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, and Holden Comprehensive Cancer CenterJoshua M Madsen - The University of Iowa Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, and Holden Comprehensive Cancer CenterMitchell C Coleman - The University of Iowa Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, and Holden Comprehensive Cancer CenterMelissa L. T Teoh - The University of Iowa Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, and Holden Comprehensive Cancer CenterScott G Westphal - The University of Iowa Carver College of MedicineDouglas R Spitz - The University of Iowa Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, and Holden Comprehensive Cancer CenterRafael Radi - Faculdad de Medicina, Universidad de la Républica de Uruguay, Montevideo, UruguayFrederick E Domann - The University of Iowa Free Radical and Radiation Biology Program, Department of Radiation Oncology, Carver College of Medicine, and Holden Comprehensive Cancer Center
- Resource Type
- Journal article
- Publication Details
- Radiation research, Vol.174(3), pp.290-296
- DOI
- 10.1667/RR2235.1
- PMID
- 20726720
- PMCID
- PMC2942016
- NLM abbreviation
- Radiat Res
- ISSN
- 0033-7587
- eISSN
- 1938-5404
- Language
- English
- Date published
- 09/2010
- Academic Unit
- Pathology; Orthopedics and Rehabilitation; Surgery; Radiation Oncology
- Record Identifier
- 9984040269802771
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