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In vitro synthesized bacterial outer membrane protein is integrated into bacterial inner membranes but translocated across microsomal membranes

Nature volume 323, pages 71–73 (1986)Cite this article

Abstract

The LamB protein is an integral membrane protein of the outer membrane of Escherichia coli. We have now found that, when synthesized in an E. coli cell-free translation system supplemented with inverted vesicles derived from the E. coli inner membrane, LamB protein is integrated into the vesicle membrane as assayed by its resistance to extraction at alkaline pH. These data suggest that the inner membrane is the primary site for integration of LamB protein prior to subsequent sorting to the outer membrane. When synthesized in a wheat germ cell-free translation system supplemented with canine microsomal membranes, LamB protein is glycosylated at one or two cryptic sites, and surprisingly, it is translocated across instead of being integrated into the vesicle membrane. We suggest that the translocation machinery of the microsomal membrane, although able to recognize the signal sequences) of LamB, is unable to recognize its stop-transfer sequences), thereby yielding translocation instead of integration.

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Author notes

  1. John F. Hunt

    Present address: Department of Molecular Biophysics and Biochemistry, Yale University, New Haven, Connecticut, 06511, USA

Authors and Affiliations

  1. Laboratory of Cell Biology, Rockefeller University, New York, New York, 10021, USA

    Makoto Watanabe, John F. Hunt & Günter Blobel

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  1. Makoto Watanabe
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  2. John F. Hunt
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  3. Günter Blobel
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Watanabe, M., Hunt, J. & Blobel, G. In vitro synthesized bacterial outer membrane protein is integrated into bacterial inner membranes but translocated across microsomal membranes. Nature 323, 71–73 (1986). https://doi.org/10.1038/323071a0

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  • DOI: https://doi.org/10.1038/323071a0

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