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

Methods in Enzymology
Anton P Le BrunJeremy H Lakey

Abstract

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 structura...Continue Reading

Citations

Aug 6, 2016·Proceedings of the National Academy of Sciences of the United States of America·Wanatchaporn ArunmaneeJeremy H Lakey
Oct 14, 2016·Cellular and Molecular Life Sciences : CMLS·Xin MengRobert C Ford
Oct 3, 2020·Biochemical Society Transactions·Nicolò ParaciniJeremy H Lakey

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