The Study of (Plant) Ion Channels Reconstituted in Planar Lipid Bilayers

摘要: The development of the patch-clamp technique by Neher and Sakmann (1976) opened door to study single ion channels. Initially performed on animal cells, 8 years later was successfully applied plant cells as well (Moran et al. 1984). One reason for delay in applying is presence a cell wall which has be removed before protoplast can patch-clamped (Takeda 1985). Today, an almost standard laboratories around world specializing electrophysiology. Despite loss wall, during experiment channel studied its more or less native membrane environment. An alternative strategy channels method based reconstitution isolated, purified artificial system, such liposome planar lipid bilayer. Obviously, these techniques are especially useful cases particular inaccessible pipette (Miller 1983). small size liposomes high turnover rate channels, however, limit use studies mediated transport performance electrophysiological measurements requires either enlargement suitable patch-clamping (Tank Miller 1983) into so-called bilayer (PLB). PLB allows level under precise, defined experimental conditions been appplied succesfully originating from membranes endomembranes (see references Labarca Latorre 1992; Coronado 1992). White Tester (1992) were first reconstitute plasma PLB. At same time, Klughammer (1992a,b) vacuolar membrane. Here we describe methodology set up system show obtained after incorporation interested reader referred superb textbooks Hanke Schlue (1993) (1986) (1992).