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A single copy of SecYEG is sufficient for preprotein translocation.

Abstract

The heterotrimeric SecYEG complex comprises a protein-conducting channel in the bacterial cytoplasmic membrane. SecYEG functions together with the motor protein SecA in preprotein translocation. Here, we have addressed the functional oligomeric state of SecYEG when actively engaged in preprotein translocation. We reconstituted functional SecYEG complexes labelled with fluorescent markers into giant unilamellar vesicles at a natively low density. Förster's resonance energy transfer and fluorescence (cross-) correlation spectroscopy with single-molecule sensitivity allowed for independent observations of the SecYEG and preprotein dynamics, as well as complex formation. In the presence of ATP and SecA up to 80% of the SecYEG complexes were loaded with a preprotein translocation intermediate. Neither the interaction with SecA nor preprotein translocation resulted in the formation of SecYEG oligomers, whereas such oligomers can be detected when enforced by crosslinking. These data imply that the SecYEG monomer is sufficient to form a functional translocon in the lipid membrane.

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  • Authors

    Kedrov A, Kusters I, Krasnikov VV, Driessen AJ

    Source

    The EMBO journal 30:21 2011 Nov 2 pg 4387-97

    MeSH

    Escherichia coli
    Escherichia coli Proteins
    Gene Dosage
    Membrane Lipids
    Membrane Proteins
    Methanococcus
    Models, Biological
    Organisms, Genetically Modified
    Protein Multimerization
    Protein Precursors
    Protein Transport
    Unilamellar Liposomes

    Pub Type(s)

    Journal Article
    Research Support, Non-U.S. Gov't

    Language

    eng

    PubMed ID

    21897368