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

    Year: 2014

    Journal: BioNanoScience June 2014, Volume 4, Issue 2, pp 104-110, 20141015

    Authors: Angélique Coutable (1)(2)(3)(4)(5), Irina Randrianjatovo (1)(2)(3), Vincent Noireaux (6), Christophe Vieu (4)(5), Christophe Thibault (4)(5), Emmanuelle Trévisiol (1)(2)(3)(4)(5), Jean M. François (1)(2)(3)

    Last authors: Jean M. François

    Organizations: 1. INSA,UPS,INP; LISBP, Université de Toulouse, 135 Avenue de Rangueil, 31077, Toulouse, France 2. INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, 31400, Toulouse, France 3. CNRS, UMR5504, 31400, Toulouse, France 4. CNRS; LAAS, 7 Avenue du Colonel Roche, 31400, Toulouse, France 5. UPS, INSA, INP, ISAE; UT1, UTM, LAAS, Université de Toulouse, 31400, Toulouse, France 6. Physics Department, University of Minnesota, Minneapolis, MN, USA

    Country: USA, United States of America, France

    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 fromStaphylococcus. 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 theStaphylococcus 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.