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Separation of liquid phases in giant vesicles of ternary mixtures of phospholipids and cholesterol
Authors:Veatch Sarah L  Keller Sarah L
Institution:Departments of Chemistry and Physics, University of Washington, Seattle, Washington 98195-1700, USA.
Abstract:We use fluorescence microscopy to directly observe liquid phases in giant unilamellar vesicles. We find that a long list of ternary mixtures of high melting temperature (saturated) lipids, low melting temperature (usually unsaturated) lipids, and cholesterol produce liquid domains. For one model mixture in particular, DPPC/DOPC/Chol, we have mapped phase boundaries for the full ternary system. For this mixture we observe two coexisting liquid phases over a wide range of lipid composition and temperature, with one phase rich in the unsaturated lipid and the other rich in the saturated lipid and cholesterol. We find a simple relationship between chain melting temperature and miscibility transition temperature that holds for both phosphatidylcholine and sphingomyelin lipids. We experimentally cross miscibility boundaries both by changing temperature and by the depletion of cholesterol with beta-cyclodextrin. Liquid domains in vesicles exhibit interesting behavior: they collide and coalesce, can finger into stripes, and can bulge out of the vesicle. To date, we have not observed macroscopic separation of liquid phases in only binary lipid mixtures.
Keywords:GUV  giant unilamellar vesicle  FRET  fluorescence resonance energy transfer  Chol  cholesterol  PC  phosphatidylcholine  DCPC  dicaprylphosphatidylcholine (di(10:0)PC)  DLPC  dilauroylphosphatidylcholine (di(12:0)PC)  DMPC  dimyristoylphosphatidylcholine (di(14:0)PC)  DPPC  dipalmitoylphosphatidylcholine (di(16:0)PC)  DSPC  distearoylphosphatidylcholine (di(18:0)PC)  DAPC  diarachidoylphosphatidylcholine (di(20:0)PC)  DOPC  dioleoylphosphatidylcholine (di(18:1)PC)  POPC  1-palmitoyl  2-oleoylphosphatidylcholine (16:0/18:1 PC)  SM  sphingomyelin  BSM  brain sphingomyelin  ESM  egg sphingomyelin  PSM  d-sphingomyelin (16:0 SM)" target="_blank">N-palmitoyl-d-sphingomyelin (16:0 SM)  SSM  d-sphingomyelin (18:0 SM)" target="_blank">N-stearoyl-d-sphingomyelin (18:0 SM)
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