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J. Biol. Chem., Vol. 280, Issue 27, 25659-25664, July 8, 2005

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Interactions of MotX with MotY and with the PomA/PomB Sodium Ion Channel Complex of the Vibrio alginolyticus Polar Flagellum^*

Mayuko Okabe, Toshiharu Yakushi, and Michio Homma

From the Division of Biological Science, Graduate School of Science, Nagoya University, Chikusa-Ku, Nagoya 464-8602, Japan

Rotation of the sodium ion-driven polar flagellum of Vibrio alginolyticus requires the inner membrane sodium ion channel complex PomA/PomB and the outer membrane components MotX and MotY. None of the detergents used in this study were able to solubilize MotX when it was expressed alone. However, when co-expressed with MotY, MotX was solubilized by some detergents. The change in the solubility of MotX suggests that MotY interacts with MotX. In agreement with this, a pull-down assay showed the association of MotY with MotX. Solubilized MotX and MotY eluted in the void volume from a gel-filtration column, suggesting that MotX and MotY form a large oligomeric structure(s). In the absence of MotY, MotX affected membrane localization of the PomA/PomB complex and of PomB alone but not of PomA alone, suggesting an interaction between MotX and PomB. We propose that MotX exhibits multiple interactions with the other motor components, first with MotY for its localization to the outer membrane and then with the PomA/PomB complex through PomB for the motor rotation.

Received for publication, January 7, 2005 , and in revised form, April 22, 2005.

^* This work was supported by grants-in-aid for scientific research from the Ministry of Education, Science, and Culture of Japan, the Japan Science and Technology Corporation (to M. H. and T. Y.), and the Soft Nano-machine Project of Japan Science and Technology Agency (to T. Y. and M. H.). 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 Biophysics and Biochemistry, Yale University, 266 Whitney Ave., Bass 225, New Haven, CT 06520-8114.

To whom correspondence should be addressed. Tel.: 81-52-789-2992; Fax: 81-52-789-3001; E-mail: 4juji{at}cc.nagoya-u.ac.jp.

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