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1.2 Å X-ray Structure of the Renal Potassium Channel Kv1.3 T1 Domain

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Abstract

Here we present the structure of the T1 domain derived from the voltage-dependent potassium channel Kv1.3 of Homo sapiens sapiens at 1.2 Å resolution crystallized under near-physiological conditions. The crystals were grown without precipitant in 150 mM KPi, pH 6.25. The crystals show I4 symmetry typical of the natural occurring tetrameric assembly of the single subunits. The obtained structural model is based on the highest resolution currently achieved for tetramerization domains of voltage-gated potassium channels. We identified an identical fold of the monomer but inside the tetramer the single monomers show a significant rotation which leads to a different orientation of the tetramer compared to other known structures. Such a rotational movement inside the tetrameric assembly might influence the gating properties of the channel. In addition we see two distinct side chain configurations for amino acids located in the top layer proximal to the membrane (Tyr109, Arg116, Ser129, Glu140, Met142, Arg146), and amino acids in the bottom layer of the T1-domain distal from the membrane (Val55, Ile56, Leu77, Arg86). The relative populations of these two states are ranging from 50:50 for Val55, Tyr109, Arg116, Ser129, Glu140, 60:40 for Met142, 65:35 for Arg86, 70:30 for Arg146, and 80:20 for Ile56 and Leu77. The data suggest that in solution these amino acids are involved in an equilibrium of conformational states that may be coupled to the functional states of the whole potassium channel.

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Abbreviations

NMR:

Nuclear magnetic resonance

NMM:

New minimal medium

TROSY:

Transversed relaxation optimized spectroscopy

HSQC:

Heteronuclear single quantum coherence

DSS:

4,4-dimethyl-4-silapentane-1-sulfonic acid

MR:

Molecular replacement

SLS:

Swiss light source

CCP4:

Collaborative Computational Project No. 4

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Acknowledgments

This work was supported by the European Union, the Fonds der Chemischen Industrie (FCI), and the Deutsche Forschungsgemeinschaft (SFB 699). The crystallographic experiments were performed on the X10SA beamline at the SLS, Paul Scherrer Institut, Villigen, Switzerland.

Conflict of interest

None.

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Author notes

  1. Michael Weyand

    Present address: Department of Biochemistry, University of Bayreuth, Universitätsstraße 30, 95447, Bayreuth, Germany

Authors and Affiliations

  1. Department of Biophysics and Physical Biochemistry, Centre of Magnetic Resonance in Chemistry and Biomedicine, University of Regensburg, Universitätsstraße 31, 93053, Regensburg, Germany

    Werner Kremer, Andreas Winklmeier, Christina Schreier & Hans Robert Kalbitzer

  2. Department of Structural Biology, Max-Planck-Institute for Molecular Physiology, Otto-Hahn-Str. 11, 44227, Dortmund, Germany

    Michael Weyand

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  1. Werner Kremer
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Corresponding authors

Correspondence to Werner Kremer or Hans Robert Kalbitzer.

Additional information

The coordinates and structure factors have been deposited in the Protein Data Bank under accession number 4BGC.

Werner Kremer and Michael Weyand have contributed equally.

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Kremer, W., Weyand, M., Winklmeier, A. et al. 1.2 Å X-ray Structure of the Renal Potassium Channel Kv1.3 T1 Domain. Protein J 32, 533–542 (2013). https://doi.org/10.1007/s10930-013-9513-2

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