Issue 3, 2021

Development, structure and mechanics of a synthetic E. coli outer membrane model

Abstract

The outer membrane (OM) of Gram-negative bacteria is a complex asymmetric bilayer containing lipids, lipopolysaccharides (LPS) and proteins. While it is a mechanical and chemical barrier, it is also the primary surface of bacterial recognition processes that involve infection by and of the bacterium. Uncovering the mechanisms of these biological functions has been hampered by the lack of suitable model systems. Here we report the step-by-step assembly of a synthetic OM model from its fundamental components. To enable the efficient formation of a supported lipid bilayer at room temperature, dimyristoyl-phosphocholine (DMPC) was used as the lipid component to which we progressively added LPS and OM proteins. The assembled system enabled us to explore the contribution of the molecular components to the topographical structure and stability of the OM. We found that LPS prefers solid-state membrane regions and forms stable vesicles in the presence of divalent cations. LPS can gradually separate from DMPC membranes to form independent vesicles, pointing at the dynamic nature of the lipid-LPS system. The addition of OM proteins from E. coli and saturating levels of LPS to DMPC liposomes resulted in a thicker and more stable bilayer the surface of which displayed a nanoscale texture formed of parallel, curved, long (>500 nm) stripes spaced apart with a 15 nm periodicity. The synthetic membrane may facilitate the investigation of binding and recognition processes on the surface of Gram-negative bacteria.

Graphical abstract: Development, structure and mechanics of a synthetic E. coli outer membrane model

Supplementary files

Article information

Article type
Paper
Submitted
24 nov 2020
Accepted
09 dec 2020
First published
16 dec 2020
This article is Open Access
Creative Commons BY license

Nanoscale Adv., 2021,3, 755-766

Development, structure and mechanics of a synthetic E. coli outer membrane model

B. Kiss, T. Bozó, D. Mudra, H. Tordai, L. Herényi and M. Kellermayer, Nanoscale Adv., 2021, 3, 755 DOI: 10.1039/D0NA00977F

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