Electrophysiological characterization of bacterial pore-forming proteins in planar lipid bilayers

Abstract

Together with patch-clamp, the planar lipid bilayer technique is one of the electrophysiological approaches used to study the biophysical properties of bacterial pore-forming proteins. Electrophysiological studies have provided important insight into the mechanistic details underlying the function of this class of proteins. Although there are different apparatus designs and variations to the process of obtaining channel recordings, the general architecture of a planar lipid bilayer setup involves two compartments filled with an ionic solution and separated by a septum with a micro-aperture, where a phospholipid bilayer is formed, and an amplifier used to clamp the membrane potential and record currents. Bacterial outer membrane porins and translocons, among others, can be reconstituted in this bilayer and their electrophysiology probed in different physicochemical conditions or through functional assays with substrates or potential modulators. This chapter describes specifically the reconstitution of detergent purified outer membrane pore-forming proteins into artificial lipid membranes using a laboratory customized planar lipid bilayer apparatus and the subsequent recording of channel activity under voltage clamp. © Springer Science+Business Media New York 2013.