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

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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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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).

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Authors and Affiliations

  1. INSA,UPS,INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France

    Angélique Coutable, Irina Randrianjatovo, Emmanuelle Trévisiol & Jean M. François

  2. INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, 31400, Toulouse, France

    Angélique Coutable, Irina Randrianjatovo, Emmanuelle Trévisiol & Jean M. François

  3. CNRS, UMR5504, 31400, Toulouse, France

    Angélique Coutable, Irina Randrianjatovo, Emmanuelle Trévisiol & Jean M. François

  4. CNRS; LAAS, 7 Avenue du Colonel Roche, 31400, Toulouse, France

    Angélique Coutable, Christophe Vieu, Christophe Thibault & Emmanuelle Trévisiol

  5. UPS, INSA, INP, ISAE; UT1, UTM, LAAS, Université de Toulouse, 31400, Toulouse, France

    Angélique Coutable, Christophe Vieu, Christophe Thibault & Emmanuelle Trévisiol

  6. Physics Department, University of Minnesota, Minneapolis, MN, USA

    Vincent Noireaux

Authors
  1. Angélique Coutable
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  2. Irina Randrianjatovo
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  3. Vincent Noireaux
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  4. Christophe Vieu
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  7. Jean M. François
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Correspondence to Angélique Coutable.

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Coutable, A., Randrianjatovo, I., Noireaux, V. et al. A Comparative Study of α-Hemolysin Expression in Supported Lipid Bilayers of Synthetic and Enriched Complex Bacterial Lipid. BioNanoSci. 4, 104–110 (2014). https://doi.org/10.1007/s12668-014-0127-8

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  • DOI: https://doi.org/10.1007/s12668-014-0127-8

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