Formation and Characterization of Supported Lipid Bilayers Composed
of Phosphatidylethanolamine and Phosphatidylglycerol by Vesicle Fusion,
a Simple but Relevant Model for Bacterial Membranes
Version 3 2019-06-26, 14:05
Version 2 2019-06-20, 13:52
Version 1 2019-06-19, 18:12
Posted on 2019-06-26 - 14:05
Supported
lipid bilayers (SLBs) are simple and robust biomimics with controlled
lipid composition that are widely used as models of both mammalian
and bacterial membranes. However, the lipids typically used for SLB
formation poorly resemble those of bacterial cell membranes due to
the lack of available protocols to form SLBs using mixtures of lipids
relevant for bacteria such as phosphatidylethanolamine (PE) and phosphatidylglycerol
(PG). Although a few reports have been published recently on the formation
of SLBs from Escherichia coli lipid
extracts, a detailed understanding of these systems is challenging
due to the complexity of the lipid composition in such natural extracts.
Here, we present for the first time a simple and reliable protocol
optimized to form high-quality SLBs using mixtures of PE and PG at
compositions relevant for Gram-negative membranes. We show using neutron
reflection and quartz microbalance not only that Ca2+ ions
and temperature are key parameters for successful bilayer deposition
but also that mass transfer to the surface is a limiting factor. Continuous
flow of the lipid suspension is thus crucial for obtaining full SLB
coverage. We furthermore characterize the resulting bilayers and report
structural parameters, for the first time for PE and PG mixtures,
which are in good agreement with those reported earlier for pure POPE
vesicles. With this protocol in place, more suitable and reproducible
studies can be conducted to understand biomolecular processes occurring
at cell membranes, for example, for testing specificities and to unravel
the mechanism of interaction of antimicrobial peptides.
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Kjellerup Lind, Tania; W. A. Skoda, Maximilian; Cárdenas, Marité (2019). Formation and Characterization of Supported Lipid Bilayers Composed
of Phosphatidylethanolamine and Phosphatidylglycerol by Vesicle Fusion,
a Simple but Relevant Model for Bacterial Membranes. ACS Publications. Collection. https://doi.org/10.1021/acsomega.9b01075
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