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Interactions between lipids and voltage sensor paddles detected with tarantula toxins

Nature Structural & Molecular Biology volume 16, pages 1080–1085 (2009)Cite this article

Abstract

Voltage-activated ion channels open and close in response to changes in voltage, a property that is essential for generating nerve impulses. Studies on voltage-activated potassium (Kv) channels show that voltage-sensor activation is sensitive to the composition of lipids in the surrounding membrane. Here we explore the interaction of lipids with S1–S4 voltage-sensing domains and find that the conversion of the membrane lipid sphingomyelin to ceramide-1-phosphate alters voltage-sensor activation in an S1–S4 voltage-sensing protein lacking an associated pore domain, and that the S3b–S4 paddle motif determines the effects of lipid modification on Kv channels. Using tarantula toxins that bind to paddle motifs within the membrane, we identify mutations in the paddle motif that weaken toxin binding by disrupting lipid-paddle interactions. Our results suggest that lipids bind to voltage-sensing domains and demonstrate that the pharmacological sensitivities of voltage-activated ion channels are influenced by the surrounding lipid membrane.

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Figure 1: Membrane modification alters gating of voltage-activated ion channels and a voltage-activated phosphatase.
Figure 2: Membrane modification alters the apparent affinity of tarantula toxins for Kv channels without altering membrane partitioning.
Figure 3: Comparison of the effects of Kv2.1 paddle mutations on GxTx-1E affinity before and after membrane modification with SMaseD.
Figure 4: Coupling energies mapped onto the X-ray structure of a Kv channel (PDB 2R9R)41 and a model for how lipid modification alters toxin affinity.
Figure 5: Model illustrating toxin binding to lipid-associated paddle motifs.

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Acknowledgements

We thank M. Holmgren, J. Kumar, M. Mayer, J. Mindell, S. Silberberg and members of the Swartz laboratory for helpful discussions and the US National Institute of Neurological Disorders and Stroke (NINDS) DNA sequencing facility for DNA sequencing. We thank Y. Okamura (Okazaki Center for Integrative Biosciences) for providing Ci-VSP complementary DNA, Y. Xu and Z. Lu (University of Pennsylvania) for supplying recombinant SMaseD and W. Schmalhofer and M. Garcia (Merck Research Labs) for supplying 125I-GxTx-1E. This work was supported by the Intramural Research Program of the NINDS, National Insitutes of Health (NIH) (to K.J.S.) and by an NIH-FWO postdoctoral fellowship (to F.B.).

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Author notes

  1. AbdulRasheed A Alabi

    Present address: Present address: Department of Molecular and Cellular Physiology, Stanford University, Stanford, California, USA.,

Authors and Affiliations

  1. Molecular Physiology and Biophysics Section, Porter Neuroscience Research Center, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, Maryland, USA

    Mirela Milescu, Frank Bosmans, AbdulRasheed A Alabi & Kenton J Swartz

  2. Laboratory of Toxicology, University of Leuven, Leuven, Belgium

    Frank Bosmans

  3. Department of Life Sciences, Gwangju Institute of Science and Technology, Gwangju, Korea

    Seungkyu Lee & Jae Il Kim

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Contributions

M.M., F.B. and A.A.A. performed molecular biology and electrophysiology experiments, and S.L. synthesized GxTx-1E. All authors contributed to the study design and to writing the manuscript.

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Correspondence to Mirela Milescu or Kenton J Swartz.

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Milescu, M., Bosmans, F., Lee, S. et al. Interactions between lipids and voltage sensor paddles detected with tarantula toxins. Nat Struct Mol Biol 16, 1080–1085 (2009). https://doi.org/10.1038/nsmb.1679

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