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J. Biol. Chem., Vol. 279, Issue 16, 16246-16253, April 16, 2004

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Gene and Protein Characterization of the Human Glutathione S-Transferase Kappa and Evidence for a Peroxisomal Localization^*

Fabrice Morel, Claudine Rauch, Elise Petit, Amélie Piton, Nathalie Theret, Brian Coles¶, and André Guillouzo

From the INSERM U456, Université de Rennes I, 2 Avenue du Professeur Léon Bernard, 35043 Rennes, France and the ^¶Division of Molecular Epidemiology, National Center for Toxicological Research, Jefferson, Arkansas 72079

Kappa class glutathione S-transferase (GST) cDNA sequences have been identified in rat, mouse, and human. In the present study, we determined the structure and chromosomal location of the human GST Kappa 1 (hGSTK1) gene, characterized the protein, and demonstrated its subcellular localization. The human gene spans 5 kb, has 8 exons, and maps onto chromosome 7q34. The 5'-flanking region lacks TATA or CCAAT boxes, but there is an initiator element overlapping the transcription start site. hGSTK1 amino acid sequence showed homology to bacterial 2-hydroxychromene-2-carboxylate isomerase, an enzyme involved in naphthalene degradation pathway. hGSTK1 mRNA was expressed in all of the organs examined. Subcellular fractionation of HepG2 cells showed that the protein was located in peroxisomes and mitochondria and was not detectable in cytoplasm. The peroxisomal localization was confirmed by transfection of HepG2 cells with a plasmid coding a green fluorescent protein fused inframe to the N terminus of hGSTK1. The C terminus of hGSTK1 was essential for localization of the protein to peroxisomes, and the C-terminal sequence Ala-Arg-Leu represents a peroxisome targeting signal. This is the first time that a human GST has been found in peroxisomes, suggesting a new function for this family of enzymes.

Received for publication, December 8, 2003 , and in revised form, January 21, 2004.

^* This work was supported in part by the Institut National de la Santé et de la Recherche Médicale and the Association pour la Recherche sur le Cancer. 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.: 33-223-23-48-10; Fax: 33-223-23-47-94; E-mail: Fabrice.Morel{at}rennes.inserm.fr.

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