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J. Biol. Chem., Vol. 279, Issue 49, 50662-50669, December 3, 2004

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Similarities between the Abiotic Reduction of Selenite with Glutathione and the Dissimilatory Reaction Mediated by Rhodospirillum rubrum and Escherichia coli^*

Janine Kessi and Kurt W. Hanselmann

From the Microbial Ecology Group, Institute of Plant Biology, University of Zürich, Zollikerstrasse 107, Zurich CH-8008, Switzerland

Various mechanisms have been proposed to explain the biological dissimilatory reduction of selenite () to elemental selenium (Se°), although none is without controversy. Glutathione, the most abundant thiol in the eukaryotic cells, the cyanobacteria, and the , , and groups of the proteobacteria, has long been suspected to be involved in selenium metabolism. Experiments with the phototrophic proteobacterium Rhodospirillum rubrum showed that the rate of selenite reduction was decreased when bacteria synthesized lower than normal levels of glutathione, and in Rhodobacter sphaeroides and Escherichia coli the reaction was reported to induce glutathione reductase. In the latter organism superoxide dismutase was also induced in cells grown in the presence of selenite, indicating that superoxide anions (O^-₂) were produced. These observations led us to investigate the abiotic (chemical) reduction of selenite by glutathione and to compare the features of this reaction with those of the reaction mediated by R. rubrum and E. coli. Our findings imply that selenite was first reduced to selenodiglutathione, which reached its maximum concentration within the 1st min of the reaction. Formation of selenodiglutathione was paralleled by a rapid reduction of cytochrome c, a known oxidant for superoxide anions. Cytochrome c reduction was inhibited by superoxide dismutase, indicating that O^-₂ was the source of electrons for the reduction. These results demonstrated that superoxide was produced in the abiotic reduction of selenite with glutathione, thus lending support to the hypothesis that glutathione may be involved in the reaction mediated by R. rubrum and E. coli. The second phase of the reaction, which led to the formation of elemental selenium (Se°), developed more slowly. Se° precipitation reached a maximum within 2 h after the beginning of the reaction. Secondary reactions leading to the degradation of the superoxide significantly decreased the yield of Se° in the abiotic reaction compared with that of the bacterially mediated selenite reduction. Abiotically formed selenium particles showed the same characteristic orange-red color, spherical structure, and size as particles produced by R. rubrum, again providing support for the hypothesis that glutathione is involved in the reduction of selenite to elemental selenium in this organism.

Received for publication, May 26, 2004 , and in revised form, August 20, 2004.

^* 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. Tel.: 41-01-634-8211; Fax: 41-01-634-8204; E-mail: Janine.kessi{at}access.unizh.ch.

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