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J Biol Chem, Vol. 274, Issue 9, 5362-5369, February 26, 1999

Individual Subunits Contribute Independently to Slow Gating of Bovine EAG Potassium Channels

Roland SchönherrDagger , Solveig HehlDagger , Heinrich Terlau§, Arnd Baumann, and Stefan H. HeinemannDagger

From the Dagger  Max-Planck-Gesellschaft, Arbeitsgruppe Molekulare und zelluläre Biophysik, Drackendorfer Strasse 1, D-07747 Jena, § Max-Planck-Institut für experimentelle Medizin, Abteilung Molekulare Biologie neuronaler Signale, Hermann-Rein-Strasse 3, D-37075 Göttingen, and  Institut für Biologische Informationsverarbeitung, Forschungszentrum Jülich, D-52425 Jülich, Germany

The bovine ether à go-go gene encodes a delayed rectifier potassium channel. In contrast to other delayed rectifiers, its activation kinetics is largely determined by the holding potential and the concentration of extracellular Mg2+, giving rise to slowly activating currents with a characteristic sigmoidal rising phase. Replacement of a single amino acid in the extracellular linker between transmembrane segments S3 and S4 (L322H) strongly reduced the prepulse dependence and accelerated activation by 1 order of magnitude. In addition, compared with the wild type, the half-activation voltage of this mutant was shifted by more than 30 mV to more negative potentials. We used dimeric and tetrameric constructs of the bovine eag1 gene to analyze channels with defined stoichiometry of mutated and wild-type subunits within the tetrameric channel complexes. With increasing numbers of mutated subunits, the channel activation was progressively accelerated, and the sigmoidicity of the current traces was reduced. Based on a quantitative analysis, we show that the slow gating, typical for EAG channels, is mediated by independent conformational transitions of individual subunits, which gain their voltage dependence from the S4 segment. At a given voltage, external Mg2+ increases the probability of a channel subunit to be in the slowly activating conformation, whereas mutation L322H strongly reduces this probability.

Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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