Interactions of N-terminal and C-terminal parts of the small conductance Ca2+ activated K+ channel, hSK3.
Ca(2+) activated K(+) channels modulate the afterhyperpolarization in neurons. Using a variety of different techniques we obtained information about the function of N- and C-terminal parts of the Ca(2+)-activated K(+) channel, SK3. By means of the yeast two hybrid technique we found an interaction between N-C and N-N- terminal parts of SK3. The strong N-C and N-N interaction was specific for SK3 and could not be observed for SK1 and SK2. Possibly a homotetrameric assembly of SK3 is favored in tissues were all SK channels are expressed. In addition, the interaction in SK3 was independent of the length of the polymorphic glutamine repeat in the N-terminus of SK3. Electrophysiological investigations showed that expression of amino acids 1-299 of SK3 (SK3N_299) modified the 1-EBIO pharmacology of endogenous SK3 channels in PC12 cells without affecting the Ca(2+)-sensitvity. The activation by 0.5 mM 1-EBIO in cells expressing amino acids 1-299 of SK3 was reduced by 32% in comparison to control experiments. Considering the N-C interaction in yeast, we conclude that the sensitivity of SK3 channels to 1-EBIO was modified by N-C interactions with SK3N_299. Therefore we conclude that N-C interactions influence SK3 channel function.
Frei
Eva
E
Department of Applied Physiology, University of Ulm, Albert-Einstein-Allee 11, D-89081 Ulm, Germany.
Spindler
Isabell
I
Grissmer
Stephan
S
Jäger
Heike
H
D000595:Amino Acid Sequence; D000818:Animals; D001562:Benzimidazoles / Q000494:pharmacology*; D002118:Calcium / Q000494:pharmacology; D002120:Calcium Channel Agonists / Q000494:pharmacology*; D004594:Electrophysiology; D016716:PC12 Cells; D024681:Potassium Channels, Calcium-Activated / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D051640:Small-Conductance Calcium-Activated Potassium Channels / Q000819:agonists / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D001562:Benzimidazoles; D002120:Calcium Channel Agonists; C496901:Kcnn3 protein, rat; D024681:Potassium Channels, Calcium-Activated; D051640:Small-Conductance Calcium-Activated Potassium Channels; C102635:1-ethyl-2-benzimidazolinone; D002118:Calcium