Amino terminal-dependent gating of the potassium channel rat eag is compensated by a mutation in the S4 segment.
1. Rat eag potassium channels (r-eag) were expressed in Xenopus oocytes. They gave rise to delayed rectifying K+ currents with a strong Cole-Moore effect. 2. Deletions in the N-terminal structure of r-eag either shifted the activation threshold to more negative potentials and slowed the activation kinetics (delta 2-190, delta 2-12 and delta 7-12) or resulted in a shift to more positive potentials and faster activation kinetics (delta 150-162). 3. The impact of the deletion delta 7-12 was investigated in more detail: it almost abolished the Cole-Moore effect and markedly slowed down channel deactivation. 4. Unlike wild-type channels, the deletion mutants delta 7-12 exhibited a rapid inactivation which, in combination with the slow deactivation, resulted in current characteristics which were similar to those of the related potassium channel HERG. 5. Both the slowing of deactivation and the inactivation induced by the deletion delta 7-12 were compensated by a single histidine-to-arginine change in the S4 segment, while this mutation (H343R) only had minor effects on the gating kinetics of the full-length r-eag channel. 6. These results demonstrate a functional role of the N-terminus in the voltage-dependent gating of potassium channels which is presumably mediated by an interaction of the N-terminal protein structure with the S4 motif during the gating process.
Terlau
H
H
Max-Planck-Institut für experimentelle Medizin, Abteilung Molekulare Biologie neuronaler Signale, Göttingen, Germany. terlau@mail.mpiem.gwdg.de
Heinemann
S H
SH
Stühmer
W
W
Pongs
O
O
Ludwig
J
J
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D051638:Ether-A-Go-Go Potassium Channels; D005260:Female; D017353:Gene Deletion; D015640:Ion Channel Gating / Q000502:physiology*; D007700:Kinetics; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000502:physiology; D009865:Oocytes / Q000502:physiology; D018408:Patch-Clamp Techniques; D015221:Potassium Channels / Q000737:chemistry / Q000235:genetics*; D017434:Protein Structure, Tertiary; D051381:Rats; D014982:Xenopus laevis
D051638:Ether-A-Go-Go Potassium Channels; D015221:Potassium Channels