Current recordings of ion channel proteins immobilized on resin beads.
Current ion channel current measurement techniques are cumbersome, as they require many steps and much time. This is especially true when reconstituting channels into liposomes and incorporating them into lipid bilayers. Here, we report a novel method that measures ion channel current more efficiently than current methods. We applied our method to KcsA and MthK channels by binding them to cobalt affinity gel beads with histidine tags and then forming a lipid bilayer membrane on the bead. This allowed channels to incorporate into the bilayer and channel currents to be measured quickly and easily. The efficiency was such that currents could be recorded with extremely low amounts of protein. In addition, the channel direction could be determined by the histidine tag. This method has the potential to be applied to various channel proteins and channel research in general.
Hirano
Minako
M
Network Center for Molecular and System Life Sciences, Graduate School of Frontier Biosciences, Osaka University, 1-3 Yamadaoka Suita, Osaka 565-0871, Japan.
Takeuchi
Yuko
Y
Aoki
Takaaki
T
Yanagida
Toshio
T
Ide
Toru
T
D002462:Cell Membrane / Q000378:metabolism; D004553:Electric Conductivity; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism*; D007473:Ion Channels / Q000737:chemistry* / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism; D008863:Microspheres; D015221:Potassium Channels / Q000737:chemistry / Q000378:metabolism
D055767:Immobilized Proteins; D007473:Ion Channels; D008051:Lipid Bilayers; D008081:Liposomes; D015221:Potassium Channels