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J. Biol. Chem., Vol. 281, Issue 47, 35667-35676, November 24, 2006

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A Derivative of Lipid A Is Involved in Signal Recognition Particle/SecYEG-dependent and -independent Membrane Integrations^*

Ken-ichi Nishiyama¹, Ayao Ikegami, Michael Moser, Emile Schiltz^¶, Hajime Tokuda, and Matthias Müller²

From the Institute of Biochemistry and Molecular Biology, Centre of Biochemistry and Molecular Cell Research, University of Freiburg, Hermann-Herder-Strasse 7, D-79104 Freiburg, Germany, Institute of Molecular and Cellular Biosciences, University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-0032, Japan, and ^¶Institute of Organic Chemistry and Biochemistry, University of Freiburg, Albertstrasse 21, D-79104 Freiburg, Germany

A cell-free system was developed that allows the correct integration of single and multispanning membrane proteins of Escherichia coli into proteoliposomes. We found that physiological levels of diacylglycerol were required to prevent spontaneous integration into liposomes even of the polytopic mannitol permease. Using diacylglycerol-containing proteoliposomes, we identified a novel integration-stimulating factor. Integration of mannitol permease was dependent on both the SecYEG translocon and this factor and was mediated by signal recognition particle and signal recognition particle receptor. Integration of M13 procoat, which is independent of both signal recognition particle/signal recognition particle receptor and SecYEG, was also promoted by this factor. Furthermore, the factor stimulated the post-translational translocation of presecretory proteins, suggesting that it also mediates integration of a signal sequence. This factor was found to be a lipid A-derived membrane component possessing a peptide moiety.

Received for publication, August 28, 2006 , and in revised form, September 22, 2006.

^* This work was supported by an EMBO long term fellowship (to K. N.), Deutsche Forschungsgemeinschaft Sonderforschungsbereich Grant 388, European Union Grant LSHG-CT-2004-005257 (to M. M.), and Ministry of Education, Science, Sports and Culture of Japan grants (to K. N. and H. T.). 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.

¹ To whom correspondence may be addressed. Tel.: 81-3-5841-7831; Fax: 81-3-5841-8464; E-mail: unishiy{at}mail.ecc.u-tokyo.ac.jp. ² To whom correspondence may be addressed. Tel.: 49-761-203-5265; Fax: 49-761-203-5274; E-mail: matthias.mueller{at}biochemie.uni-freiburg.de.

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