Hybrid potassium channels by tandem linkage of inactivating and non-inactivating subunits.
We constructed tandem cDNA by linking the 5' end of a delayed rectifier-type (Kv1.2) clone to the 3' end of a transient-type (Kv1.4) K+ channel clone. Fusion genes were also constructed, consisting of Kv1.4 and mutants of Kv1.2, which have a single amino acid substitution in the S4-S5 loop. From electrophysiological characterization, it is likely that two pairs of tandem heterodimer constructs can form hybrid channels. In addition, it has been revealed that the wild-type hybrid channel shows a time constant of inactivation very similar to that observed in the homotetrameric Kv1.4 channel. Difference of inactivation kinetics between wild-type and mutant hybrid K+ channels suggests that not only the S4-S5 loop of Kv1.4 but also that of Kv1.2 can serve as the acceptor sites for the inactivation gates, and that all of four sets of loops should be functional for rapid inactivation. From these results, in the hybrid channels the structure and composition of the acceptor sites could be important factors for determining the rate of inactivation.
Nunoki
K
K
Department of Pharmacology, Tohoku University School of Medicine, Sendai, Japan.
Ishii
K
K
Okada
H
H
Yamagishi
T
T
Murakoshi
H
H
Taira
N
N
D000595:Amino Acid Sequence; D000818:Animals; D018076:DNA, Complementary; D007700:Kinetics; D008564:Membrane Potentials; D008969:Molecular Sequence Data; D026902:Potassium Channel Blockers; D015221:Potassium Channels / Q000235:genetics* / Q000502:physiology; D011993:Recombinant Fusion Proteins; D014981:Xenopus
D018076:DNA, Complementary; D026902:Potassium Channel Blockers; D015221:Potassium Channels; D011993:Recombinant Fusion Proteins