A Comparative Study of α-Hemolysin Expression in Supported Lipid Bilayers of Synthetic and Enriched Complex Bacterial Lipid

Angélique Coutable; Irina Randrianjatovo; Vincent Noireaux; Christophe Vieu; Christophe Thibault; Emmanuelle Trévisiol; Jean M. François

doi:10.1007/s12668-014-0127-8

A Comparative Study of α-Hemolysin Expression in Supported Lipid Bilayers of Synthetic and Enriched Complex Bacterial Lipid

Angélique Coutable, Irina Randrianjatovo, Vincent Noireaux, Christophe Vieu, Christophe Thibault, Emmanuelle Trévisiol, Jean M. François

Physics and Astronomy (Twin Cities)

Research output: Contribution to journal › Article › peer-review

1 Scopus citations

Abstract

The purpose of this short communication was to examine whether the formation of a supported lipid bilayer (SLB) made with purely synthetic lipid or with Escherichia coli complex lipid may have any influence on the production and incorporation of the transmembrane protein α-hemolysin from Staphylococcus. Different molar ratio of E. coli total extract lipid and synthetic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were used to prepare SLB, which were characterized by combination of quartz crystal microbalance with dissipation monitoring (QCM-D), fluorescence recovery after photobleaching (FRAP), and atomic force microscopy (AFM). It was found that a 68:32 molar ratio was optimal to produce a SLB mimicking a bacterial lipid membrane. Comparing this later SLB with a purely synthetic SLB (100 % POPC) as receptacle for expression of the Staphylococcus α-hemolysin fused to eGFP by a cell-free expression system (CFES), we showed that both production and incorporation of this membrane protein was very similar.

Original language	English (US)
Pages (from-to)	104-110
Number of pages	7
Journal	BioNanoScience
Volume	4
Issue number	2
DOIs	https://doi.org/10.1007/s12668-014-0127-8
State	Published - Jun 2014

Bibliographical note

Funding Information:
Acknowledgments This work was supported by the French National Research Agency (ANR) (FLANAMOVE project). The content of this work is the sole responsibility of the authors. QCM-D experiments and AFM imaging were performed at the “Institut des Technologies Avancées en Sciences du Vivant” (Toulouse, France).

Keywords

Biomimetic lipid bilayer
Cell-free transcription-translation
Membrane protein
Supported lipid bilayer (SLB)
α-Hemolysin

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This output contributes to the following UN Sustainable Development Goals (SDGs)

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title = "A Comparative Study of α-Hemolysin Expression in Supported Lipid Bilayers of Synthetic and Enriched Complex Bacterial Lipid",

abstract = "The purpose of this short communication was to examine whether the formation of a supported lipid bilayer (SLB) made with purely synthetic lipid or with Escherichia coli complex lipid may have any influence on the production and incorporation of the transmembrane protein α-hemolysin from Staphylococcus. Different molar ratio of E. coli total extract lipid and synthetic 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were used to prepare SLB, which were characterized by combination of quartz crystal microbalance with dissipation monitoring (QCM-D), fluorescence recovery after photobleaching (FRAP), and atomic force microscopy (AFM). It was found that a 68:32 molar ratio was optimal to produce a SLB mimicking a bacterial lipid membrane. Comparing this later SLB with a purely synthetic SLB (100 % POPC) as receptacle for expression of the Staphylococcus α-hemolysin fused to eGFP by a cell-free expression system (CFES), we showed that both production and incorporation of this membrane protein was very similar.",

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A Comparative Study of α-Hemolysin Expression in Supported Lipid Bilayers of Synthetic and Enriched Complex Bacterial Lipid

Abstract

Bibliographical note

Keywords

UN SDGs

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