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J. Biol. Chem., Vol. 278, Issue 27, 25179-25190, July 4, 2003
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From the
The Jackson Laboratory, Bar Harbor, Maine 04609, the Department of Biology, Fairfield University, Fairfield, Connecticut 06430, and the ¶AstraZeneca R&D Molndal, S-43183 Molndal, Sweden
Reactive oxygen species, especially hydrogen peroxide, are important in cellular signal transduction. However, excessive amounts of these species damage tissues and cells by oxidizing virtually all important biomolecules. Peroxiredoxin 6 (PRDX6) (also called antioxidant protein 2, or AOP2) is a novel peroxiredoxin family member whose function in vivo is unknown. Through immunohistochemistry, we have determined that the PRDX6 protein was widely expressed in every tissue examined, most abundantly in epithelial cells. It was found in cytosol, but not in membranes, organelles, and nuclei fractions. Prdx6 mRNA was also expressed in every tissue examined. The widespread expression of Prdx6 suggested that its functions were quite important. To determine these functions, we generated Prdx6-targeted mutant (Prdx6–/–) mice, confirmed the gene disruption by Southern blots, PCR, RT-PCR, Western blots, and immunohistochemistry, and compared the effects of paraquat, hydrogen peroxide, and t-butyl hydroperoxide on Prdx6–/– and wild-type (Prdx6+/+) macrophages, and of paraquat on Prdx6–/– and Prdx6+/+ mice. Prdx6–/– macrophages had higher hydrogen peroxide levels, and lower survival rates; Prdx6–/– mice had significantly lower survival rates, more severe tissue damage, and higher protein oxidation levels. Additionally, there were no differences in the mRNA expression levels of other peroxiredoxins, glutathione peroxidases, catalase, superoxide dismutases, thioredoxins, and glutaredoxins between normal Prdx6–/– and Prdx6+/+ mice and those injected with paraquat. Our study provides in vivo evidence that PRDX6 is a unique non-redundant antioxidant that functions independently of other peroxiredoxins and antioxidant proteins.
Received for publication, March 17, 2003 , and in revised form, April 28, 2003.
* This study was supported by AstraZeneca, Sweden and the Core Grant CA34196 from the National Institutes of Health. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
|| To whom correspondence should be addressed: The Jackson Laboratory, 600 Main St., Bar Harbor, ME 04609. Tel.: 207-288-6388; Fax: 207-288-6078; E-mail: bjp{at}jax.org.
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