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

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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 languageEnglish (US)
Pages (from-to)104-110
Number of pages7
JournalBioNanoScience
Volume4
Issue number2
DOIs
StatePublished - 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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