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Oxidative Stress Response in an Anaerobic Hyperthermophilic Archaeon: Presence of a Functional Peroxiredoxin in Pyrococcus horikoshii
- Source :
- Journal of Biochemistry. 136:541-547
- Publication Year :
- 2004
- Publisher :
- Oxford University Press (OUP), 2004.
-
Abstract
- Oxidative stress response in an anaerobic hyperthermophilic archaeon Pyrococcus horikoshii OT-3 was analyzed by two-dimensional gel electrophoresis. When P. horikoshii was grown on medium supplemented with air, a marked increase in the level of a 25-kDa protein was observed in comparison with cells grown under anaerobic conditions. The N-terminal amino acid sequence of the protein was determined to be VVIGEKFPEVEVKTTHGVIKLPDYF, which coincides with that of the putative alkyl hydroperoxide reductase that has been predicted in the genome database of P. horikoshii. The gene (PH1217) encoding the protein was cloned and expressed in Escherichia coli. The produced enzyme was a hyperthermostable peroxiredoxin whose activity was not lost after incubation at 90 degrees C for 20 min. The enzyme catalyzes the reduction of cumene hydroperoxide and hydrogen peroxide using dithiothreitol as an electron donor. Northern blot analysis revealed that the transcription of the gene increased by the addition of exogenous oxygen and by the addition of an oxidative stress-inducing reagent, and reached maximum within 30 min. These results suggest that the peroxiredoxin plays an important role in the peroxide-scavenging system in an anaerobic archaeon P. horikoshii.
- Subjects :
- Hot Temperature
Time Factors
Archaeal Proteins
Molecular Sequence Data
Electrons
Oxidative phosphorylation
Biology
Reductase
medicine.disease_cause
Biochemistry
Dithiothreitol
chemistry.chemical_compound
Pyrococcus horikoshii
Databases, Genetic
Escherichia coli
medicine
Humans
Electrophoresis, Gel, Two-Dimensional
Amino Acid Sequence
Cloning, Molecular
Molecular Biology
Genome
Sequence Homology, Amino Acid
Peroxiredoxins
General Medicine
Hydrogen-Ion Concentration
Blotting, Northern
biology.organism_classification
Molecular biology
Recombinant Proteins
Hyperthermophile
Culture Media
Protein Structure, Tertiary
Oxygen
Oxidative Stress
Models, Chemical
Peroxidases
chemistry
Cumene hydroperoxide
RNA
Electrophoresis, Polyacrylamide Gel
Peroxiredoxin
Subjects
Details
- ISSN :
- 0021924X
- Volume :
- 136
- Database :
- OpenAIRE
- Journal :
- Journal of Biochemistry
- Accession number :
- edsair.doi.dedup.....82f17fe63fdc899739e50250cf347e81