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Asymmetric phospholipid: lipopolysaccharide bilayers; a Gram-negative bacterial outer membrane mimic

Lookup NU author(s): Anton Le Brun, Professor Jeremy LakeyORCiD

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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.


Publication metadata

Author(s): Clifton LA, Skoda MWA, Daulton EL, Hughes AV, Le Brun AP, Lakey JH, Holt SA

Publication type: Article

Publication status: Published

Journal: Journal of the Royal Society Interface

Year: 2013

Volume: 10

Issue: 89

Print publication date: 16/10/2013

Date deposited: 02/05/2014

ISSN (print): 1742-5689

ISSN (electronic): 1742-5662

Publisher: Royal Society Publishing

URL: http://dx.doi.org/10.1098/rsif.2013.0810

DOI: 10.1098/​rsif.2013.0810


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Funding

Funder referenceFunder name
1310101ISIS beam-time award
080342Wellcome Trust
093581Wellcome Trust

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