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Originally published In Press as doi:10.1074/jbc.M007216200 on January 2, 2001

J. Biol. Chem., Vol. 276, Issue 13, 10407-10412, March 30, 2001
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Effect of Oxygen on Induction of the Cystine Transporter by Bacterial Lipopolysaccharide in Mouse Peritoneal Macrophages*

Hideyo SatoDagger §, Kazumi Kuriyama-MatsumuraDagger §, Taro HashimotoDagger , Hiromi SasakiDagger , Hongyu WangDagger , Tetsuro IshiiDagger , Giovanni E. Mann||, and Shiro BannaiDagger

From the Dagger  Department of Biochemistry, Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575 Japan and the || Centre for Cardiovascular Biology and Medicine, Guy's, King's, and St. Thomas' School of Biomedical Sciences, King's College London, LONDON SE1 1UL, United Kingdom

Amino acid transport in mouse peritoneal macrophages is mediated by several membrane carriers with different substrate specificity and sensitivity to environmental stimuli. We reported previously that transport activities of cystine and arginine in the macrophages were induced markedly by low concentrations of bacterial lipopolysaccharide (LPS). It is known that a variety of macrophage functions are affected by ambient oxygen tension. In this study, we have investigated the effects of oxygen on the induction of amino acid transport activity by LPS and found that the induction of cystine, but not arginine, transport activity was dependent on the ambient oxygen tension. When the macrophages were cultured with 2% O2 in the presence of 1 ng/ml LPS, induction of cystine transport activity was reduced by ~70% compared with cells cultured under normoxic conditions. In macrophages, transport of cystine is mediated by a Na+-independent anionic amino acid transporter named system x<UP><SUB><RM><IT>c</IT></RM></SUB><SUP><RM><IT>−</IT></RM></SUP></UP>. System x<UP><SUB><RM><IT>c</IT></RM></SUB><SUP><RM><IT>−</IT></RM></SUP></UP> is composed of two protein components, xCT and 4F2hc, and the expression of xCT was closely correlated with system x<UP><SUB><RM><IT>c</IT></RM></SUB><SUP><RM><IT>−</IT></RM></SUP></UP> activity. A putative NF-kappa B binding site was found in the 5'-flanking region of the xCT gene, but the enhanced expression of xCT by LPS and oxygen was not mediated by NF-kappa B binding. An increase in intracellular GSH in macrophages paralleled induction of xCT, but not gamma -glutamylcysteine synthetase. These results suggest the importance of system x<UP><SUB><RM><IT>c</IT></RM></SUB><SUP><RM><IT>−</IT></RM></SUP></UP> in antioxidant defense in macrophages exposed to LPS and oxidative stress.

* This work was supported by the British Council and Japanese Ministry of Education, Science, and Technology.The costs of publication of this article were defrayed in part by the payment of page charges. The article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

The nucleotide sequence(s) reported in this paper has been submitted to the GenBankTM/EMBL Data Bank with accession number(s) AB037650-AB037661.

§ These two authors contributed equally to this work.

To whom correspondence should be addressed. Tel.: 81-298-53-3282; Fax: 81-298-53-3039; E-mail: hideyo-s@md.tsukuba.ac.jp.

Copyright © 2001 by The American Society for Biochemistry and Molecular Biology, Inc.
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