title: Versatile in vitro system to study translocation and functional integration of bacterial outer membrane proteins.
abstract: Gram-negative bacteria use the type-V secretion pathway to expose proteins at their cell surface, many of which have virulence functions. Translocation of those proteins across the outer membrane occurs either by means of dedicated translocator proteins (two-partner secretion) or covalently fused translocator domains (autotransporters). Translocator proteins and translocator domains are β-barrels requiring the β-barrel assembly machinery (BAM) for membrane integration. However, the molecular details of their passage across the envelope and insertion into the outer membrane remain enigmatic, owing in part to the fact that in vitro systems are not available. Here we describe a versatile in vitro reconstitution system that faithfully reproduces both branches of the type-V secretion pathway and the assembly of β-barrel outer membrane proteins. This system will allow an in-depth analysis of protein secretion across and integration into outer membranes.
authors.affiliation: 1] Institute of Biochemistry and Molecular Biology, Zentrum für Biochemie und Molekulare Zellforschung, University of Freiburg, Freiburg D-79104, Germany [2] Faculty of Biology, University of Freiburg, Freiburg D-79104, Germany.
authors.forename: Derrick
authors.initials: D
authors.lastname: Norell
authors.affiliation: Research Institute of Molecular Pathology, Vienna A-1030, Austria.
authors.forename: Alexander
authors.initials: A
authors.lastname: Heuck
authors.affiliation: Institute of Biochemistry and Molecular Biology, Zentrum für Biochemie und Molekulare Zellforschung, University of Freiburg, Freiburg D-79104, Germany.
authors.forename: Thuy-Anh
authors.initials: TA
authors.lastname: Tran-Thi
authors.affiliation: Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, Stockholm SE-10691, Sweden.
authors.forename: Hansjörg
authors.initials: H
authors.lastname: Götzke
authors.affiliation: Center for Infection and Immunity of Lille, Institut Pasteur de Lille, Lille F-59019, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Jacob-Dubuisson
authors.affiliation: Research Institute of Molecular Pathology, Vienna A-1030, Austria.
authors.forename: Tim
authors.initials: T
authors.lastname: Clausen
authors.affiliation: Center for Biomembrane Research, Department of Biochemistry and Biophysics, Stockholm University, Stockholm SE-10691, Sweden.
authors.forename: Daniel O
authors.initials: DO
authors.lastname: Daley
authors.affiliation: Department of Protein Evolution, Max Planck Institute for Developmental Biology, Tübingen D-72076, Germany.
authors.forename: Volkmar
authors.initials: V
authors.lastname: Braun
authors.affiliation: Institute of Biochemistry and Molecular Biology, Zentrum für Biochemie und Molekulare Zellforschung, University of Freiburg, Freiburg D-79104, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Müller
authors.affiliation: Institute of Biochemistry and Molecular Biology, Zentrum für Biochemie und Molekulare Zellforschung, University of Freiburg, Freiburg D-79104, Germany.
authors.forename: Enguo
authors.initials: E
authors.lastname: Fan
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000378:metabolism*; D002474:Cell-Free System; D011510:Proteolipids; D000069320:Type V Secretion Systems
chemical_list: D001425:Bacterial Outer Membrane Proteins; D011510:Proteolipids; D000069320:Type V Secretion Systems; C020485:proteoliposomes