Reovirus FAST Proteins Drive Pore Formation and Syncytiogenesis Using a Novel Helix-Loop-Helix Fusion-Inducing Lipid Packing Sensor
Fig 5
Partitioning of p15HPpep into liposomes is based on curvature-dependent hydrophobic defects.
(A) Partitioning curves of bis-ANS in 100 mM suspensions of 50 nm, 100 nm, and 400 nm liposomes. bis-ANS fluorescence was fit to one-site binding hyperbola nonlinear regression using GraphPad Prism. Data points represent the mean of three experiments, with error bars contained within the symbols on the graph. (B) HPLC chromatograms in arbitrary units of absorbance at 215 nm of the top (liposomes) and bottom (free peptide) fractions from sucrose gradients following partitioning of wt p15HPpep into liposomes of the indicated diameter as described in Fig 4A, except liposomes were pre-treated with 10 mM bis-ANS before p15HPpep addition. Note that retention times of the wt p15HPpep differ from Fig 4 due to use of a different HPLC column. (C) As in panel B, quantifying percent p15HPpep associated with top (black bars) and bottom (white bars) sucrose fractions based on integrating the area under the chromatogram peaks. Results are mean SEM from three independent experiments.