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Interactions between Sec complex and prepro-alpha-factor during posttranslational protein transport into the endoplasmic reticulum.

Kathrin Plath·Barrie M Wilkinson·Colin J Stirling·Tom A Rapoport
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Abstract

Posttranslational translocation of prepro-alpha-factor (ppalphaF) across the yeast endoplasmic reticulum membrane begins with the binding of the signal sequence to the Sec complex, a membrane component consisting of the trimeric Sec61p complex and the tetrameric Sec62p/63p complex. We show by photo-cross-linking that the signal sequence is bound directly to a site where it contacts simultaneously Sec61p and Sec62p, suggesting that there is a single signal sequence recognition step. We found no evidence for the simultaneous contact of the signal sequence with two Sec61p molecules. To identify transmembrane segments of Sec61p that line the actual translocation pore, a late translocation intermediate of ppalphaF was generated with photoreactive probes incorporated into the mature portion of the polypeptide. Cross-linking to multiple regions of Sec61p was observed. In contrast to the signal sequence, neighboring positions of the mature portion of ppalphaF had similar interactions with Sec61p. These data suggest that the channel pore is lined by several transmembrane segments, which have no significant affinity for the translocating polypeptide chain.

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Biological TransportCell CompartmentationCloning, MolecularEndoplasmic ReticulumIntracellular MembranesMating FactorMembrane ProteinsMembrane Transport ProteinsMutationPeptidesProtein BindingProtein BiosynthesisProtein PrecursorsProtein Processing, Post-TranslationalProtein Sorting SignalsProtein Structure, TertiarySEC Translocation ChannelsSaccharomyces cerevisiaeSaccharomyces cerevisiae ProteinsJournal ArticleResearch Support, Non-U.S. Gov'tResearch Support, U.S. Gov't, P.H.S.

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Kathrin Plath, Barrie M Wilkinson, Colin J Stirling, Tom A Rapoport. 2003-11-14. Interactions between Sec complex and prepro-alpha-factor during posttranslational protein transport into the endoplasmic reticulum.. https://doi.org/10.1091/mbc.e03-06-0390

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