title: Low resolution structure and dynamics of a colicin-receptor complex determined by neutron scattering.
abstract: Proteins that translocate across cell membranes need to overcome a significant hydrophobic barrier. This is usually accomplished via specialized protein complexes, which provide a polar transmembrane pore. Exceptions to this include bacterial toxins, which insert into and cross the lipid bilayer itself. We are studying the mechanism by which large antibacterial proteins enter Escherichia coli via specific outer membrane proteins. Here we describe the use of neutron scattering to investigate the interaction of colicin N with its outer membrane receptor protein OmpF. The positions of lipids, colicin N, and OmpF were separately resolved within complex structures by the use of selective deuteration. Neutron reflectivity showed, in real time, that OmpF mediates the insertion of colicin N into lipid monolayers. This data were complemented by Brewster Angle Microscopy images, which showed a lateral association of OmpF in the presence of colicin N. Small angle neutron scattering experiments then defined the three-dimensional structure of the colicin N-OmpF complex. This revealed that colicin N unfolds and binds to the OmpF-lipid interface. The implications of this unfolding step for colicin translocation across membranes are discussed.
authors.affiliation: ISIS Spallation Neutron Source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0QX, United Kingdom.
authors.forename: Luke A
authors.initials: LA
authors.lastname: Clifton
authors.affiliation: Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom.
authors.forename: Christopher L
authors.initials: CL
authors.lastname: Johnson
authors.affiliation: Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom.
authors.forename: Alexandra S
authors.initials: AS
authors.lastname: Solovyova
authors.affiliation: Partnership for Structural Biology, Institut Laue Langevin, 6 Rue Jules Horowitz, 38042 Grenoble, France.
authors.forename: Phil
authors.initials: P
authors.lastname: Callow
authors.affiliation: Center for Structural Molecular Biology, Oak Ridge National Laboratory, Oak Ridge, Tennessee 37831.
authors.forename: Kevin L
authors.initials: KL
authors.lastname: Weiss
authors.affiliation: Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom.
authors.forename: Helen
authors.initials: H
authors.lastname: Ridley
authors.affiliation: Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom.
authors.forename: Anton P
authors.initials: AP
authors.lastname: Le Brun
authors.affiliation: ISIS Spallation Neutron Source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0QX, United Kingdom.
authors.forename: Christian J
authors.initials: CJ
authors.lastname: Kinane
authors.affiliation: ISIS Spallation Neutron Source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0QX, United Kingdom.
authors.forename: John R P
authors.initials: JRP
authors.lastname: Webster
authors.affiliation: ISIS Spallation Neutron Source, Rutherford Appleton Laboratory, Harwell Science and Innovation Campus, Didcot, Oxfordshire OX11 0QX, United Kingdom.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Holt
authors.affiliation: Institute for Cell and Molecular Biosciences, The Medical School, University of Newcastle, Framlington Place, Newcastle-upon-Tyne NE2 4HH, United Kingdom. Electronic address: Jeremy.Lakey@ncl.ac.uk.
authors.forename: Jeremy H
authors.initials: JH
authors.lastname: Lakey
mesh_terms: D003087:Colicins / Q000737:chemistry* / Q000378:metabolism*; D003902:Detergents / Q000737:chemistry; D004926:Escherichia coli / Q000166:cytology / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D008958:Models, Molecular; D033363:Neutron Diffraction; D010715:Phosphatidylglycerols / Q000378:metabolism; D018272:Porins / Q000378:metabolism*; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D053838:Scattering, Small Angle; D013499:Surface Properties; D013997:Time Factors
chemical_list: D003087:Colicins; D003902:Detergents; D029968:Escherichia coli Proteins; C109237:OmpF protein; D010715:Phosphatidylglycerols; D018272:Porins; C030345:1,2-dipalmitoylphosphatidylglycerol
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