title: Purification and functional reconstitution of the bacterial protein translocation pore, the SecYEG complex.
abstract: 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.
authors.affiliation: Department of Molecular Microbiology, Groningen Biomolecular Sciences and Biotechnology Institute and Zernike Institute for Advanced Materials, University of Groningen, Groningen, The Netherlands.
authors.forename: Ilja
authors.initials: I
authors.lastname: Kusters
authors.forename: Geert
authors.initials: G
authors.lastname: van den Bogaart
authors.forename: Janny
authors.initials: J
authors.lastname: de Wit
authors.forename: Viktor
authors.initials: V
authors.lastname: Krasnikov
authors.forename: Bert
authors.initials: B
authors.lastname: Poolman
authors.forename: Arnold
authors.initials: A
authors.lastname: Driessen
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000378:metabolism*; D002479:Inclusion Bodies / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D011498:Protein Precursors / Q000378:metabolism; D000069816:SEC Translocation Channels
chemical_list: D001425:Bacterial Outer Membrane Proteins; D029968:Escherichia coli Proteins; D008081:Liposomes; D011498:Protein Precursors; D000069816:SEC Translocation Channels; C057184:outer membrane protein A precursor (E coli)