Abstract
Lipid composition plays an important role in fusion of vesicles to membranes, an essential process for exocytosis. Lipid head group, tail structure, and sterol content all impact the complex phase behavior of membranes. To determine the effect of lipids on fusion rate, we utilized the nystatin/ergosterol (nys/erg) fusion assay and stimulated fusion with an osmotic gradient. Using PE/PC planar membranes with increasing concentrations of cholesterol, and keeping vesicle lipids constant (PE/PC/PS/ERG), we observed increasing fusion rates. We also observed that temperature affects fusion rates by forming DPPC membranes on 5ul glass capillary pipets. Planar membranes were painted with concentrated DPPC in decane. Significantly different fusion rates were observed at temperatures above and below the DPPC transition temperature (Tt) of 41C. Decreasing the temperature below Tt increased the fusion rate, while increasing the temperature above Tt reduced the fusion rate. These data are consistent with the hypothesis that vesicle fusion with a membrane is suppressed in a liquid disordered lipid phase, and shows how membrane fusion can be affected by lipid behavior.
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