Purification and Functional Reconstitution of the Bacterial Protein Translocation Pore, the SecYEG Complex

I. Kusters, G. van den Bogaart, J. de Wit, V. Krasnikov, B. Poolman, A. Driessen

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In bacteria, proteins are secreted across the cytoplasmic membrane by a protein complex termed translocase. The ability to study the activity of the translocase in vitro using purified proteins has been instrumental for our understanding of the mechanisms underlying this process. Here, we describe the protocols for the purification and reconstitution of the SecYEG complex in an active state into liposomes. In addition, fluorescence based in vitro assays are described that allow monitoring translocation activity discontinuously and in real time.
Original languageEnglish
Title of host publicationProtein Secretion
Subtitle of host publicationMethods and Protocols
EditorsAnastassios Economou
PublisherHumana Press
Number of pages13
ISBN (Electronic)978-1-60327-412-8
ISBN (Print)978-1-60327-167-7, 978-1-4939-5707-1
Publication statusPublished - 25-Jan-2010

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