title: Asymmetric phospholipid: lipopolysaccharide bilayers; a Gram-negative bacterial outer membrane mimic.
abstract: The Gram-negative bacterial outer membrane (OM) is a complex and highly asymmetric biological barrier but the small size of bacteria has hindered advances in in vivo examination of membrane dynamics. Thus, model OMs, amenable to physical study, are important sources of data. Here, we present data from asymmetric bilayers which emulate the OM and are formed by a simple two-step approach. The bilayers were deposited on an SiO2 surface by Langmuir-Blodgett deposition of phosphatidylcholine as the inner leaflet and, via Langmuir-Schaefer deposition, an outer leaflet of either Lipid A or Escherichia coli rough lipopolysaccharides (LPS). The membranes were examined using neutron reflectometry (NR) to examine the coverage and mixing of lipids between the bilayer leaflets. NR data showed that in all cases, the initial deposition asymmetry was mostly maintained for more than 16 h. This stability enabled the sizes of the headgroups and bilayer roughness of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine and Lipid A, Rc-LPS and Ra-LPS to be clearly resolved. The results show that rough LPS can be manipulated like phospholipids and used to fabricate advanced asymmetric bacterial membrane models using well-known bilayer deposition techniques. Such models will enable OM dynamics and interactions to be studied under in vivo-like conditions.
authors.affiliation: ISIS Pulsed Neutron and Muon Source, Science and Technology Facilities Council, Rutherford Appleton Laboratory, , Harwell, Oxfordshire OX11 OQX, UK.
authors.forename: Luke A
authors.initials: LA
authors.lastname: Clifton
authors.forename: Maximilian W A
authors.initials: MW
authors.lastname: Skoda
authors.forename: Emma L
authors.initials: EL
authors.lastname: Daulton
authors.forename: Arwel V
authors.initials: AV
authors.lastname: Hughes
authors.forename: Anton P
authors.initials: AP
authors.lastname: Le Brun
authors.forename: Jeremy H
authors.initials: JH
authors.lastname: Lakey
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Holt
mesh_terms: D002462:Cell Membrane / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism / Q000648:ultrastructure*; D006090:Gram-Negative Bacteria / Q000378:metabolism / Q000648:ultrastructure*; D008050:Lipid A / Q000737:chemistry; D008051:Lipid Bilayers / Q000737:chemistry*; D008070:Lipopolysaccharides / Q000737:chemistry*; D008567:Membranes, Artificial
chemical_list: D008050:Lipid A; D008051:Lipid Bilayers; D008070:Lipopolysaccharides; D008567:Membranes, Artificial
keywords: Gram-negative bacterial outer membrane; isotopic labelling; lipopolysaccharide; neutron reflection; rough mutant lipopolysaccharides