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Related Experiment Video

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Experimental Methods for Trapping Ions Using Microfabricated Surface Ion Traps
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Published on: August 17, 2017

Intermediate state trapping of a voltage sensor.

Jérôme J Lacroix1, Stephan A Pless, Luca Maragliano

  • 1Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, IL 60637, USA.

The Journal of General Physiology
|November 28, 2012
PubMed
Summary
This summary is machine-generated.

Mutations in the Shaker Kv channel

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Area of Science:

  • Molecular and Cellular Neuroscience
  • Biophysics
  • Structural Biology

Background:

  • Voltage-gated ion channels (VGICs) control cellular excitability via voltage sensor domains (VSDs).
  • The precise molecular mechanisms governing VSD conformational transitions remain incompletely understood.
  • Understanding VSD dynamics is crucial for deciphering ion channel function and regulation.

Purpose of the Study:

  • To investigate the role of specific mutations in the Shaker Kv channel's S1 segment on VSD movement and gating.
  • To elucidate the molecular basis for VSD stabilization in intermediate conformations.
  • To explore the contribution of non-natural amino acids and interactions to VSD function.

Main Methods:

  • Site-directed mutagenesis of the Shaker Kv channel (I241, F244, R2).
  • Electrophysiology to record voltage dependence and ionic conductance.
  • Computational modeling (string method) to determine VSD transition pathways.
  • In silico mutagenesis and molecular dynamics simulations.

Main Results:

  • I241W mutation immobilized the VSD in an intermediate state, significantly shifting voltage activation.
  • Charge-conserving R2K mutation partially rescued the I241W phenotype, while R2Q did not.
  • F244W mutation mimicked key aspects of the I241W phenotype.
  • Indole nitrogen, not cation-π interactions, was critical for the I241W effect.
  • Computational analysis suggested a stabilizing hydrogen bond involving the indole nitrogen of I241W and R2 backbone.

Conclusions:

  • Specific mutations can stabilize intermediate VSD conformations in voltage-gated ion channels.
  • A hydrogen bond interaction likely underlies the observed stabilization by the I241W mutation.
  • This study provides novel insights into VSD gating mechanisms and intermediate states using a minimally perturbing approach.