Functional analysis of Kv1.2 and paddle chimera Kv channels in planar lipid bilayers.
Voltage-dependent K(+) (Kv) channels play key roles in shaping electrical signaling in both excitable and nonexcitable cells. These channels open and close in response to the voltage changes across the cell membrane. Many studies have been carried out in order to understand the voltage-sensing mechanism. Our laboratory recently determined the atomic structures of a mammalian Kv channel Kv1.2 and a mutant of Kv1.2 named the 'paddle chimera' channel, in which the voltage sensor paddle was transferred from Kv2.1 to Kv1.2. These two structures provide atomic descriptions of voltage-dependent channels with unprecedented clarity. Until now, the functional integrity of these two channels biosynthesized in yeast cells has not been assessed. Here, we report the electrophysiological and pharmacological properties of Kv1.2 and the paddle chimera channels in planar lipid bilayers. We demonstrate that Pichia yeast produce 'normally functioning' mammalian Kv channels with qualitatively similar features to the Shaker K(+) channel in the absence of the N-terminal inactivation gate and that the paddle chimera mutant channel functions as well as Kv1.2. We find, however, that in several respects, the Kv1.2 channel exhibits functional properties that are distinct from Kv1.2 channels reported in the literature.
Tao
Xiao
X
Howard Hughes Medical Institute, Department of Molecular Neurobiology and Biophysics, Rockefeller University, 1230 York Avenue, New York, NY 10065, USA.
MacKinnon
Roderick
R
D000595:Amino Acid Sequence; D000818:Animals; D018999:Charybdotoxin / Q000494:pharmacology; D015640:Ion Channel Gating / Q000187:drug effects; D051663:Kv1.2 Potassium Channel / Q000737:chemistry / Q000378:metabolism*; D008051:Lipid Bilayers / Q000378:metabolism*; D008969:Molecular Sequence Data; D010455:Peptides / Q000494:pharmacology; D017433:Protein Structure, Secondary; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism*; D016415:Sequence Alignment; D013111:Spider Venoms / Q000494:pharmacology
D051663:Kv1.2 Potassium Channel; D008051:Lipid Bilayers; D010455:Peptides; D011993:Recombinant Fusion Proteins; D013111:Spider Venoms; C487989:VSTX1 protein, Grammostola spatulata; D018999:Charybdotoxin