Yeast thioredoxin peroxidase expression enhances the resistance of Escherichia coli to oxidative stress induced by singlet oxygen.
Singlet oxygen ((1)O(2)) is a highly reactive form of molecular oxygen that may harm living systems by oxidizing critical cellular macromolecules. A soluble protein from Saccharomyces cerevisiae specifically provides protection against a thiol-containing metal-catalyzed oxidation system (thiol/Fe(3+)/O(2)) but not against an oxidation system without thiol. This 25 kDa protein acts as a peroxidase but requires the NADPH-dependent thioredoxin system or a thiol-containing intermediate, and was named thioredoxin peroxidase (TPx). The role of TPx in the cellular defense against oxidative stress induced by singlet oxygen was investigated in Escherichia coli containing an expression vector with a yeast genomic DNA fragment that encodes TPx and mutant in which the catalytically essential amino acid cysteine (Cys-47) has been replaced with alanine by a site-directed mutagenesis. Upon exposure to methylene blue and visible light, which generates singlet oxygen, there was a distinct difference between the two strains in regard to growth kinetics, viability, the accumulation of oxidized proteins and lipids, and modulation of activities of superoxide dismutase and catalase. The results suggest that TPx may play an important protective role in a singlet oxygen-mediated cellular damage.
Kim
Sun Yee
SY
Department of Biochemistry, College of Natural Sciences, Kyungpook National University, Taegu 702-701, Korea.
Kim
Eun Ju
EJ
Park
Jeen-Woo
JW
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics* / Q000502:physiology; D007700:Kinetics; D009363:Neoplasm Proteins; D018384:Oxidative Stress / Q000502:physiology*; D010544:Peroxidases / Q000235:genetics / Q000378:metabolism*; D054464:Peroxiredoxins; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D026082:Singlet Oxygen / Q000502:physiology*
D009363:Neoplasm Proteins; D026082:Singlet Oxygen; D010544:Peroxidases; D054464:Peroxiredoxins