A mutation in S6 of Shaker potassium channels decreases the K+ affinity of an ion binding site revealing ion-ion interactions in the pore.
Under physiological conditions, potassium channels are extraordinarily selective for potassium over other ions. However, in the absence of potassium, certain potassium channels can conduct sodium. Sodium flux is blocked by the addition of low concentrations of potassium. Potassium affinity, and therefore the ability to block sodium current, varies among potassium channel subtypes (Korn, S.J., and S.R. Ikeda. 1995. Science. 269:410-412; Starkus, J.G., L. Kuschel, M.D. Rayner, and S.H. Heinemann. 1997. J. Gen. Physiol. 110:539-550). The Shaker potassium channel conducts sodium poorly in the presence of very low (micromolar) potassium due to its high potassium affinity (Starkus, J.G., L. Kuschel, M.D. Rayner, and S.H. Heinemann. 1997. J. Gen. Physiol. 110:539-550; Ogielska, E.M., and R. W. Aldrich. 1997. Biophys. J. 72:A233 [Abstr.]). We show that changing a single residue in S6, A463C, decreases the apparent internal potassium affinity of the Shaker channel pore from the micromolar to the millimolar range, as determined from the ability of potassium to block the sodium currents. Independent evidence that A463C decreases the apparent affinity of a binding site in the pore comes from a study of barium block of potassium currents. The A463C mutation decreases the internal barium affinity of the channel, as expected if barium blocks current by binding to a potassium site in the pore. The decrease in the apparent potassium affinity in A463C channels allows further study of possible ion interactions in the pore. Our results indicate that sodium and potassium can occupy the pore simultaneously and that multiple occupancy results in interactions between ions in the channel pore.
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University, Stanford, California 94305, USA.
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Ogielska
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Aldrich
D000595:Amino Acid Sequence; D000818:Animals; D001464:Barium / Q000494:pharmacology; D001665:Binding Sites / Q000502:physiology; D001692:Biological Transport / Q000235:genetics; D015640:Ion Channel Gating / Q000187:drug effects / Q000502:physiology*; D008969:Molecular Sequence Data; D016296:Mutagenesis / Q000502:physiology; D009865:Oocytes / Q000737:chemistry / Q000502:physiology; D018408:Patch-Clamp Techniques; D011188:Potassium / Q000493:pharmacokinetics*; D015221:Potassium Channels / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D051637:Shaker Superfamily of Potassium Channels; D012964:Sodium / Q000493:pharmacokinetics; D014982:Xenopus laevis
D015221:Potassium Channels; D011993:Recombinant Fusion Proteins; D051637:Shaker Superfamily of Potassium Channels; D001464:Barium; D012964:Sodium; D011188:Potassium