Inhibition of human ether a go-go potassium channels by Ca(2+)/calmodulin.

Intracellular Ca(2+) inhibits voltage-gated potassium channels of the ether a go-go (EAG) family. To identify the underlying molecular mechanism, we expressed the human version hEAG1 in Xenopus oocytes. The channels lost Ca(2+) sensitivity when measured in cell-free membrane patches. However, Ca(2+) sensitivity could be restored by application of recombinant calmodulin ...
(CaM). In the presence of CaM, half inhibition of hEAG1 channels was obtained in 100 nM Ca(2+). Overlay assays using labelled CaM and glutathione S-transferase (GST) fusion fragments of hEAG1 demonstrated direct binding of CaM to a C-terminal domain (hEAG1 amino acids 673-770). Point mutations within this section revealed a novel CaM-binding domain putatively forming an amphipathic helix with both sides being important for binding. The binding of CaM to hEAG1 is, in contrast to Ca(2+)-activated potassium channels, Ca(2+) dependent, with an apparent K(D) of 480 nM. Co-expression experiments of wild-type and mutant channels revealed that the binding of one CaM molecule per channel complex is sufficient for channel inhibition.
Mesh Terms:
Amino Acid Sequence, Animals, Binding Sites, Calcium, Calmodulin, Cation Transport Proteins, DNA-Binding Proteins, Ether-A-Go-Go Potassium Channels, Humans, Ion Channel Gating, Molecular Sequence Data, Potassium Channel Blockers, Potassium Channels, Potassium Channels, Voltage-Gated, Protein Binding, Recombinant Proteins, Sequence Homology, Amino Acid, Trans-Activators, Xenopus laevis
EMBO J.
Date: Jul. 03, 2000
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