Electrophysiological Characterization of Transport Across Outer-Membrane Channels from Gram-Negative Bacteria in Presence of Lipopolysaccharides.
Angew Chem Int Ed Engl
; 59(22): 8517-8521, 2020 05 25.
Article
em En
| MEDLINE
| ID: mdl-32023354
ABSTRACT
Multi-drug resistance in Gram-negative bacteria is often associated with low permeability of the outer membrane. To investigate the role of membrane channels in the uptake of antibiotics, we present an approach using fusion of native outer membrane vesicles (OMVs) into a planar lipid bilayer, allowing characterization of membrane protein channels in their native environment. Two major membrane channels from E. coli, OmpF and OmpC, were overexpressed from the host and the corresponding OMVs were collected. Each OMV fusion surprisingly revealed only single or few channel activities. The asymmetry of the OMVs translates after fusion into the lipid membrane with the lipopolysaccharides (LPS) dominantly present at the side of OMV addition. Compared to the conventional reconstitution method, the channels fused from OMVs containing LPS have similar conductance but a much broader distribution and significantly lower permeation. We suggest using outer membrane vesicles for functional and structural studies of membrane channels in the native membrane.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Membrana Celular
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Lipopolissacarídeos
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Fenômenos Eletrofisiológicos
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Bactérias Gram-Negativas
Idioma:
En
Ano de publicação:
2020
Tipo de documento:
Article