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Originally published In Press as doi:10.1074/jbc.M110832200 on April 30, 2002

J. Biol. Chem., Vol. 277, Issue 27, 24049-24056, July 5, 2002
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Oxidation of Ikappa Balpha at Methionine 45 Is One Cause of Taurine Chloramine-induced Inhibition of NF-kappa B Activation*

Atsuhiro KanayamaDagger §, Jun-ichiro Inoue||, Yoshioko Sugita-Konishi**Dagger Dagger , Makoto ShimizuDagger , and Yusei Miyamoto§§¶¶

From the Dagger  Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo, Tokyo 113-8657, Japan, the  Department of Applied Chemistry, Faculty of Science and Technology, Keio University, 3-14-1 Hiyoshi, Kohoku, Yokohama, Kanagawa 223-8522, Japan, the ** Department of Biomedical Food Research, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku, Tokyo 162-8640, Japan, and the §§ Department of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan

A band shift of Ikappa Balpha was observed in Western blots with Jurkat cells treated with 1 mM taurine chloramine (TauCl) for 1 h. TauCl treatment inhibited tumor necrosis factor alpha  (TNFalpha )-initiated nuclear factor kappa B (NF-kappa B) activation. TauCl did not inhibit either the upstream of Ikappa B kinase (IKK) activation or IKK itself but did inhibit NF-kappa B activation induced by IKK overexpression. Deletion experiments showed that a TauCl modification site causing the band shift of Ikappa Balpha is Met45. High performance liquid chromatography and mass spectrometry analyses of a small peptide containing Met45 revealed that TauCl oxidizes Met45. A mutant of Ikappa Balpha whose Met45 was converted to alanine did not generate a band shift upon TauCl treatment and degraded in response to TNFalpha stimulation. However, a reporter assay revealed that NF-kappa B-dependent luciferase expression was not fully recovered in cells transfected with this mutant. These results indicate that Met45 oxidation of Ikappa Balpha is a molecular mechanism underlying the TauCl-induced inhibition of NF-kappa B activation. A similar band shift was observed when HL-60 cells expressing myeloperoxidase were treated with 100 µM hydrogen peroxide for 5 min. When rat neutrophils were incubated with bacteria, intracellular taurine decreased interleukin-8 production. Therefore, taurine may help suppress excessive inflammatory reaction in neutrophils.

* This work was supported in part by a grant-in-aid for Scientific Research from the Ministry of Education, Science, Sports, and Culture of Japan and a research grant from Taisho Pharmaceutical Co., Ltd. (to Y. M.). 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.

§ Present address: Dept. of Molecular Biology, NA 8.202, University of Texas Southwestern Medical Center, 600 Harry Hines Blvd., Dallas, TX 75390-9148.

|| Present address: Division of Cellular Molecular Biology, Dept. of Cancer Biology, The Inst. of Medical Science, University of Tokyo, 4-6-1 Shirogane, Minato, Tokyo 108-8639, Japan.

Dagger Dagger Present address: Division of Microbiology, National Institutes of Health Sciences, 1-18-1 Kamiyoga, Setagaya, Tokyo 158-8501, Japan.

¶¶ To whom correspondence should be addressed: Dept. of Integrated Biosciences, Graduate School of Frontier Sciences, University of Tokyo, Bioscience Bldg. 402, 5-1-5 Kashiwanoha, Kashiwa, Chiba 277-8562, Japan. Tel.: 81-471-36-3628; Fax: 81-471-36-3630; E-mail: yusei74@k.u-tokyo.ac.jp.

Copyright © 2002 by The American Society for Biochemistry and Molecular Biology, Inc.
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