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J. Biol. Chem., Vol. 280, Issue 21, 20261-20267, May 27, 2005

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Membrane Topology of the STT3 Subunit of the Oligosaccharyl Transferase Complex^*

Hyun Kim, Gunnar von Heijne, and IngMarie Nilsson

From the Department of Biochemistry and Biophysics, Stockholm University, SE-10691 Stockholm, Sweden

The highly conserved membrane protein STT3 is part of the oligosaccharyl transferase complex in the endoplasmic reticulum of eukaryotic cells. Various experimental observations strongly suggest that STT3 contains the active site of the complex. Here, we report a detailed topology study of STT3 from two different organisms, Saccharomyces cerevisiae and mouse, using in vivo and in vitro topology mapping assays. Our results suggest that STT3 has 11 transmembrane helices and an overall N_cyt-C_lum orientation.

Received for publication, October 28, 2004 , and in revised form, February 16, 2005.

^* This work was supported by grants from the Swedish Cancer Foundation (to I. N. and G. v. H.), from the Swedish Research Council (to G. v. H.), from the Swedish Foundation for International Cooperation in Research and Higher Education (to I. N.), and from Magnus Bergvalls Stiftelse (to I. N.). 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.

The on-line version of this article (available at http://www.jbc.org) contains select STT3 hits from PBLAST via Expasy.

To whom correspondence should be addressed. Tel.: 46-8-162728; Fax: 46-8-153679; E-mail: ingmarie{at}dbb.su.se.

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