title: Structural and Functional Basis for Lipid Synergy on the Activity of the Antibacterial Peptide ABC Transporter McjD.
abstract: The lipid bilayer is a dynamic environment that consists of a mixture of lipids with different properties that regulate the function of membrane proteins; these lipids are either annular, masking the protein hydrophobic surface, or specific lipids, essential for protein function. In this study, using tandem mass spectrometry, we have identified specific lipids associated with the Escherichia coli ABC transporter McjD, which translocates the antibacterial peptide MccJ25. Using non-denaturing mass spectrometry, we show that McjD in complex with MccJ25 survives the gas phase. Partial delipidation of McjD resulted in reduced ATPase activity and thermostability as shown by circular dichroism, both of which could be restored upon addition of defined E. coli lipids. We have resolved a phosphatidylglycerol lipid associated with McjD at 3.4 Å resolution, whereas molecular dynamic simulations carried out in different lipid environments assessed the binding of specific lipids to McjD. Combined, our data show a synergistic effect of zwitterionic and negatively charged lipids on the activity of McjD; the zwitterionic lipids provide structural stability to McjD, whereas the negatively charged lipids are essential for its function.
authors.affiliation: From the Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, United Kingdom.
authors.forename: Shahid
authors.initials: S
authors.lastname: Mehmood
authors.affiliation: the Centre for Molecular Simulation and Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada.
authors.forename: Valentina
authors.initials: V
authors.lastname: Corradi
authors.affiliation: the Department of Life Sciences, Imperial College London, South Kensington, London SW7 2AZ, United Kingdom.|the Membrane Protein Lab.|the Rutherford Appleton Laboratory, Research Complex at Harwell, Oxfordshire OX11 0DE, United Kingdom, and.
authors.forename: Hassanul G
authors.initials: HG
authors.lastname: Choudhury
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire, OX11 0DE, United Kingdom.
authors.forename: Rohanah
authors.initials: R
authors.lastname: Hussain
authors.affiliation: the Department of Life Sciences, Imperial College London, South Kensington, London SW7 2AZ, United Kingdom.|the Membrane Protein Lab.|the Rutherford Appleton Laboratory, Research Complex at Harwell, Oxfordshire OX11 0DE, United Kingdom, and.
authors.forename: Patrick
authors.initials: P
authors.lastname: Becker
authors.affiliation: Diamond Light Source, Harwell Science and Innovation Campus, Oxfordshire, OX11 0DE, United Kingdom.
authors.forename: Danny
authors.initials: D
authors.lastname: Axford
authors.affiliation: the Communication Molecules and Adaptation of Microorganisms Laboratory (MCAM, UMR 7245 CNRS-MNHN), Sorbonne Universités, Muséum National d'Histoire Naturelle, Centre National de la Recherche Scientifique, CP 54, 57 Rue Cuvier, 75005 Paris, France.
authors.forename: Severine
authors.initials: S
authors.lastname: Zirah
authors.affiliation: the Communication Molecules and Adaptation of Microorganisms Laboratory (MCAM, UMR 7245 CNRS-MNHN), Sorbonne Universités, Muséum National d'Histoire Naturelle, Centre National de la Recherche Scientifique, CP 54, 57 Rue Cuvier, 75005 Paris, France.
authors.forename: Sylvie
authors.initials: S
authors.lastname: Rebuffat
authors.affiliation: the Centre for Molecular Simulation and Department of Biological Sciences, University of Calgary, Calgary, Alberta T2N 1N4, Canada.
authors.forename: D Peter
authors.initials: DP
authors.lastname: Tieleman
authors.affiliation: From the Department of Chemistry, University of Oxford, South Parks Road, Oxford OX1 3QZ, United Kingdom, carol.robinson@chem.ox.ac.uk.
authors.forename: Carol V
authors.initials: CV
authors.lastname: Robinson
authors.affiliation: the Department of Life Sciences, Imperial College London, South Kensington, London SW7 2AZ, United Kingdom, kbeis@imperial.ac.uk.|the Membrane Protein Lab.|the Rutherford Appleton Laboratory, Research Complex at Harwell, Oxfordshire OX11 0DE, United Kingdom, and.
authors.forename: Konstantinos
authors.initials: K
authors.lastname: Beis
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism; D001430:Bacteriocins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D056004:Molecular Dynamics Simulation; D010715:Phosphatidylglycerols / Q000737:chemistry* / Q000378:metabolism; D000072417:Protein Domains; D013329:Structure-Activity Relationship
chemical_list: D018528:ATP-Binding Cassette Transporters; D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D029968:Escherichia coli Proteins; C000604081:McjD protein, E coli; D010715:Phosphatidylglycerols; C020411:microcin
keywords: ABC transporter; circular dichroism (CD); delipidation; lipid; membrane protein; membrane transport; molecular dynamics; non-denaturing mass spectrometry; structural biology; structure-function