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Related Experiment Videos

Voltage-dependent structural interactions in the Shaker K(+) channel.

S K Tiwari-Woodruff1, M A Lin, C T Schulteis

  • 1Department of Physiology and Molecular Biology Institute, University of California, Los Angeles, School of Medicine, Los Angeles, California 90095-1751, USA.

The Journal of General Physiology
|February 2, 2000
PubMed
Summary
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Structural interactions in Shaker K(+) channel voltage sensors were investigated. Residue E283 in S2 interacts with S4 residues R368 and R371, influencing channel gating and conformational states during activation.

Area of Science:

  • Molecular biology
  • Biophysics
  • Ion channel function

Background:

  • Shaker K(+) channels are crucial for neuronal excitability.
  • Voltage-dependent activation involves conformational changes in the voltage sensor.
  • Previous work suggested structural interactions between S2 and S4 domains.

Purpose of the Study:

  • To test the hypothesis that S4 residues R368 and R371 interact with S2 residue E283 in specific conformational states.
  • To elucidate the role of these interactions in the voltage-dependent activation pathway of Shaker K(+) channels.

Main Methods:

  • Cysteine substitution and reactivity assays to determine residue location at different potentials.
  • Double charge reversal mutations to probe interactions between specific residues.

Related Experiment Videos

  • Functional analysis of mutant channels to assess gating properties.

    • Main Results:

    • Residue E283 is located near the extracellular surface in both resting and activated states.
    • Mutations at E283/R371 stabilized an activated channel conformation, slowing transitions.
    • Mutations at E283/R368 stabilized a closed, partially activated conformation.

    Conclusions:

    • E283 interacts with R371 in activated states and with R368 in intermediate closed states.
    • These interactions are critical for the conformational rearrangements during Shaker K(+) channel activation.
    • A preliminary model for voltage sensor structural rearrangements is proposed.