Reconstituting channels into planar membranes: a conceptual framework and methods for fusing vesicles to planar bilayer phospholipid membranes.
Protocols to reconstitute channels into planar bilayers via fusion methods have now been developed. The greater the intravesicular pressures generated, the greater is the fusion. These pressures can be calculated exactly for any experimental configuration. For some of the configurations, adding nystatin channels to the vesicle membrane will greatly aid fusion. The configurations of the 1990 Method (Figs. 4 and 5) are optimal for fusing vesicles that are reconstituted with ion-selective channels to planar membranes. Greater binding, and ultimately greater fusion, is achieved by ejecting vesicles directly at the membrane rather than by simply adding material to the cis compartment.
Cohen
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Department of Physiology, Rush Medical College, Chicago, Illinois 61612.
Niles
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D000666:Amphotericin B; D004875:Ergosterol; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005559:Formamides; D007202:Indicators and Reagents; D007473:Ion Channels / Q000502:physiology*; D007700:Kinetics; D008051:Lipid Bilayers; D008561:Membrane Fusion; D008722:Methods; D008954:Models, Biological; D009761:Nystatin; D010743:Phospholipids / Q000737:chemistry*; D013816:Thermodynamics
D005452:Fluoresceins; D005456:Fluorescent Dyes; D005559:Formamides; D007202:Indicators and Reagents; D007473:Ion Channels; D008051:Lipid Bilayers; D010743:Phospholipids; D009761:Nystatin; C031066:formamide; D000666:Amphotericin B; C007740:fluorexon; D004875:Ergosterol