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Originally published In Press as doi:10.1074/jbc.M200936200 on March 20, 2002

J. Biol. Chem., Vol. 277, Issue 21, 19080-19086, May 24, 2002
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Characterization and Expression of L-Amino Acid Oxidase of Mouse Milk*

Youping SunDagger , Eriko NonobeDagger , Youko Kobayashi, Takeshi Kuraishi, Fugaku Aoki§, Kazuo Yamamoto§, and Senkiti Sakai

From the Department of Animal Breeding, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Yayoi 1-1-1, Bunkyo-ku, Tokyo 113-8657 and the § Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwa City, Chiba 277-8562, Japan

L-Amino acid oxidase (LAO) was purified from mouse milk. LAO reacted with L-amino acids in an apparent order of Phe > Met, Tyr > Cys, Leu > His other 11 amino acids tested and produced H2O2 in a dose- and time-dependent manner. LAO in milk had a molecular mass of about 113 kDa and was converted to a 60-kDa protein by SDS-PAGE. LAO consisted of two subunits. The N- and C-terminal amino acid sequence determination followed by cDNA cloning showed that the 60-kDa protein consisted of 497 amino acids. LAO mRNA spanned about 2.0 kb, and its expression was found only in the mammary epithelial cells. Glucocorticoid was essential for LAO gene expression. Thus, the LAO gene is expressed acutely upon the onset of milk synthesis. LAO mRNA increased 1 day before parturition, peaked during early to mid-lactation, and decreased at the end of lactation. This is the first demonstration showing that LAO is present in milk. Mastitis is caused by an intramammary bacterial infection. As mouse milk produced H2O2 using endogenous free amino acids, we suggest that LAO, together with free amino acids, is responsible for killing bacteria in the mammary gland.

* This work was supported by Grants-in-aid for Scientific Research 10460122 and 10660266 from the Ministry of Education, Science, Sports and Culture of Japan and by the Tikusan-Gijutu Kyokai at Tokyo.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/EBI Data Bank with accession number(s) AB034801.

Dagger Both authors contributed equally to this work.

To whom correspondence should be addressed. Tel.: 81-3-5841-5380; Fax: 81-3-5841-8180; E-mail: asenkiti@mail.ecc.u-tokyo.ac.jp.

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