title: Assessing the Outer Membrane Insertion and Folding of Multimeric Transmembrane β-Barrel Proteins.
abstract: In addition to the cytoplasmic membrane, Gram-negative bacteria have a second lipid bilayer, the outer membrane, which is the de facto barrier between the cell and the extracellular milieu. Virtually all integral proteins of the outer membrane form β-barrels, which are inserted into the outer membrane by the BAM complex. Some outer membrane proteins, like the porins and trimeric autotransporter adhesins, are multimeric. In the former case, the porin trimer consists of three individual β-barrels, whereas in the latter, the single autotransporter β-barrel domain is formed by three separate polypeptides. This chapter reviews methods to investigate the folding and membrane insertion of multimeric OMPs and further explains the use of a BamA depletion strain to study the effects of the BAM complex on multimeric OMPs in E. coli.
authors.affiliation: Department of Biosciences, University of Oslo, Oslo, 0316, Norway.
authors.forename: Jack C
authors.initials: JC
authors.lastname: Leo
authors.affiliation: Interfaculty Institute for Microbiology and Infection Medicine, University Clinics Tübingen, Tübingen, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Oberhettinger
authors.affiliation: Department of Biosciences, University of Oslo, Oslo, 0316, Norway. dirk.linke@ibv.uio.no.|Department of Protein Evolution, Max Planck Institute for Developmental Biology, Tübingen, 72076, Germany. dirk.linke@ibv.uio.no.
authors.forename: Dirk
authors.initials: D
authors.lastname: Linke
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D006358:Hot Temperature; D008958:Models, Molecular; D017510:Protein Folding; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary
chemical_list: D001425:Bacterial Outer Membrane Proteins; C503914:BamA protein, E coli; D029968:Escherichia coli Proteins
keywords: Adhesin; BAM complex; BamA depletion strain; Outer membrane; Trimeric autotransporter; β-barrel