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Deuterium Labeling Strategies for Creating Contrast in Structure-Function Studies of Model Bacterial Outer Membranes Using Neutron Reflectometry

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

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Abstract

© 2016 Elsevier B.V. All rights reserved. Studying the outer membrane of Gram-negative bacteria is challenging due to the complex nature of its structure. Therefore, simplified models are required to undertake structure-function studies of processes that occur at the outer membrane/fluid interface. Model membranes can be created by immobilizing bilayers to solid supports such as gold or silicon surfaces, or as monolayers on a liquid support where the surface pressure and fluidity of the lipids can be controlled. Both model systems are amenable to having their structure probed by neutron reflectometry, a technique that provides a one-dimensional depth profile through a membrane detailing its thickness and composition. One of the strengths of neutron scattering is the ability to use contrast matching, allowing molecules containing hydrogen and those enriched with deuterium to be highlighted or matched out against the bulk isotopic composition of the solvent. Lipopolysaccharides, a major component of the outer membrane, can be isolated for incorporation into model membranes. Here, we describe the deuteration of lipopolysaccharides from rough strains of Escherichia coli for incorporation into model outer membranes, and how the use of deuterated materials enhances structural analysis of model membranes by neutron reflectometry.


Publication metadata

Author(s): Le Brun AP, Clifton LA, Holt SA, Holden PJ, Lakey JH

Editor(s): Kumar CV

Publication type: Book Chapter

Publication status: Published

Book Title: Methods in Enzymology

Year: 2017

Volume: 566

Pages: 231-252

Print publication date: 01/05/2017

Online publication date: 18/06/2015

Acceptance date: 01/01/1900

Publisher: Academic Press Inc.

Place Published: San Diego

URL: https://doi.org/10.1016/bs.mie.2015.05.020

DOI: 10.1016/bs.mie.2015.05.020

Library holdings: Search Newcastle University Library for this item

ISBN: 9780128105023


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