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Originally published In Press as doi:10.1074/jbc.M107819200 on August 31, 2001

J. Biol. Chem., Vol. 276, Issue 45, 41576-41579, November 9, 2001
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Serum Amyloid P Component Is the Shiga Toxin 2-neutralizing Factor in Human Blood*

Tsuyoshi KimuraDagger §, Shinobu TaniDagger , Yoh-ichi Matsumoto, and Tae Takeda||

From the Dagger  Teijin Institute for Biomedical Research, Teijin Ltd., 4-3-2, Asahigaoka, Hino City, Tokyo 191-8512, Japan,  Pharmaceutical Development Department I, Teijin Ltd., 2-1-1, Uchisaiwaicho, Chiyoda-ku, Tokyo 100-8585, Japan, and || Department of Infectious Diseases Research, National Children's Medical Research Center, 3-35-31, Taishido, Setagaya-ku, Tokyo 154-8509, Japan

It has been suggested that some factor present in human plasma binds to Shiga toxin 2 (Stx2) and neutralizes it in vitro (Bitzan, M., Klemt, M., Steffens, R., and Muller-Wiefel, D. E. (1993) Infection 21, 140-145). This factor does not exist in other species (Caprioli, A., Luzzi, I., Seganti, L., Marchetti, M., Karmali, M., Clarke, I., and Boyd, B. (1994) Recent Adv. VTEC Infect. 353-356). Because analysis of this factor is important to understanding the pathology induced by Shiga toxin-producing Escherichia coli, we purified this factor from human plasma and identified it. Purification was carried out by serially subjecting human plasma to Con A-Sepharose, DEAE-Sepharose, hydroxyapatite, and gel-filtration high performance liquid chromatography (HPLC), using Stx2-neutralizing activity as the indicator. The gel-filtration HPLC fraction yielded a single band on SDS-polyacrylamide gel electrophoresis. Twenty N-terminal amino acid residues of this fraction were analyzed and found to correspond perfectly to human serum amyloid P component (HuSAP). Because commercially available HuSAP also showed Stx2 binding and neutralizing activity, we identified this factor as HuSAP.

* This work was supported by the Program for Promotion of Fundamental Studies of the Organization for Pharmaceutical Safety and Research of Japan.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.

§ To whom correspondence should be addressed. Tel.: +81-42-586-8137; Fax: +81-42-587-5515; E-mail: t.kimura@teijin.co.jp.

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