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Control of K+ channel gating by protein phosphorylation: structural switches of the inactivation gate

Nature Structural Biology volume 6, pages 146–150 (1999)Cite this article

Abstract

Fast N–type inactivation of voltage–dependent potassium (Kv) channels controls membrane excitability and signal propagation in central neurons and occurs by a 'ball–and–chain'–type mechanism. In this mechanism an N–terminal protein domain (inactivation gate) occludes the pore from the cytoplasmic side. In Kv3.4 channels, inactivation is not fixed but is dynamically regulated by protein phosphorylation. Phosphorylation of several identified serine residues on the inactivation gate leads to reduction or removal of fast inactivation. Here, we investigate the structure–function basis of this phospho–regulation with nuclear magnetic resonance (NMR) spectroscopy and patch–clamp recordings using synthetic inactivation domains (ID). The dephosphorylated ID exhibited compact structure and displayed high–affinity binding to its receptor. Phosphorylation of serine residues in the N– or C–terminal half of the ID resulted in a loss of overall structural stability. However, depending on the residue(s) phosphorylated, distinct structural elements remained stable. These structural changes correlate with the distinct changes in binding and unbinding kinetics underlying the reduced inactivation potency of phosphorylated IDs.

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Figure 1: Effect of phosphorylation on inactivation kinetics is site specific.
Figure 2: Effect of phosphorylation is not due to changes in net charge of the inactivation gate.
Figure 3: Structure of the phospho ID S–P 15,21 and the dephospho Raw3–ID.
Figure 4: Structure of the phospho ID S–P 8 and the dephospho Raw3–ID.

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Acknowledgements

The authors would like to thank K.C. Holmes for continuous support, M. Geyer for technical help and J.P. Adelman, J. Maylie, J. Mosbacher and G. Yellen for helpful comments and reading of the manuscript. The work was supported by the Deutsche Forschungsgemeinschaft (Fa 332/2–1).

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Authors and Affiliations

  1. Department of Physiology II, University of Tübingen, Ob dem Himmelreich 7, Tübingen, 72074, Germany

    Christoph Antz, Johann Peter Ruppersberg, Thomas Baukrowitz & Bernd Fakler

  2. Institute of Physiological Chemistry, University of Tübingen, Ob dem Himmelreich 7, Tübingen, 72074, Germany

    Tanja Bauer & Hubert Kalbacher

  3. Zentrum für Molekulare Biologie, Im Neuenheimer Feld, Heidelberg, 69120, Germany

    Rainer Frank

  4. Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, 1020 Locust St JAH 245, Philadelphia, 19107, Pennsylvania, USA

    Manuel Covarrubias

  5. Department of Biophysics and Physical Biochemistry, University of Regensburg, Universitätsstr. 31, Regensburg, 93053, Germany

    Hans Robert Kalbitzer

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Correspondence to Bernd Fakler.

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Antz, C., Bauer, T., Kalbacher, H. et al. Control of K+ channel gating by protein phosphorylation: structural switches of the inactivation gate. Nat Struct Mol Biol 6, 146–150 (1999). https://doi.org/10.1038/5833

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