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A sodium-mediated structural switch that controls the sensitivity of Kir channels to PtdIns(4,5)P2

Nature Chemical Biology volume 4, pages 624–631 (2008)Cite this article

Abstract

Inwardly rectifying potassium (Kir) channels are gated by the membrane phospholipid phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2). Among them, Kir3 requires additional molecules, such as the βγ subunits of G proteins or intracellular sodium, for channel gating. Using an interactive computational-experimental approach, we show that sodium sensitivity of Kir channels involves the side chains of an aspartate and a histidine located across from each other in a crucial loop in the cytosolic domain, as well as the backbone carbonyls of two more residues and a water molecule. The location of the coordination site in the vicinity of a conserved arginine shown to affect channel–PtdIns(4,5)P2 interactions suggests that sodium triggers a structural switch that frees the crucial arginine. Mutations of the aspartate and the histidine that affect sodium sensitivity also enhance the channel's sensitivity to PtdIns(4,5)P2. Furthermore, on the basis of the molecular characteristics of the coordination site, we identify and confirm experimentally a sodium-sensitive phenotype in Kir5.1.

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Figure 1: Positions of key residues in a loop of Kir channels where Na+ is coordinated.
Figure 2: H-bonding pattern between position 217 and residues in its vicinity.
Figure 3: Experimental evidence for Kir3.4* residues involved in Na+ sensitivity.
Figure 4
Figure 5: Experimental evidence for Kir2.1 and Kir5.1 residues involved in Na+ sensitivity.

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Acknowledgements

We thank E. Findeis, T. Borges, V. Petrou and H. Vaananen for oocyte preparation. This work was supported by a US National Institutes of Health grant (HL-59949) to D.E.L.

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Author notes

  1. Avia Rosenhouse-Dantsker, Jin L Sui, Radda Rusinova & Diomedes E Logothetis

    Present address: Present addresses: Department of Medicine, Section of Pulmonary, Critical Care and Sleep Medicine, University of Illinois at Chicago, 840 South Wood Street (M/C 719), Room 920-N CSB, Chicago, Illinois 60612, USA (A.R.-D.), CombinatoRx Inc., 245 First Street, Cambridge, Massachusetts 02142, USA (J.L.S.), Department of Physiology and Biophysics, Weill Medical College of Cornell University, 1300 York Avenue, New York, New York 10021, USA (R.R.) and Virginia Commonwealth University, Medical College of Virginia Campus, Department of Physiology and Biophysics, Sanger Hall 3-005, 1101 East Marshall Street, Richmond, Virginia 23298, USA (D.E.L.).,

Authors and Affiliations

  1. Department of Structural and Chemical Biology, Mount Sinai School of Medicine, One Gustave L. Levy Place, New York, 10029, New York, USA

    Avia Rosenhouse-Dantsker, Jin L Sui, Qi Zhao, Radda Rusinova & Diomedes E Logothetis

  2. Department of Physiology and Biophysics, Virginia Commonwealth University, Medical College of Virginia Campus, Sanger Hall 3-005, 1101 East Marshall Street, Richmond, 23298, Virginia, USA

    Aldo A Rodríguez-Menchaca & Zhe Zhang

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Contributions

A.R.-D. designed and performed the molecular modeling studies, the mutagenesis and the two-electrode voltage-clamp experiments, wrote the manuscript and revised the manuscript to address the reviewers' comments. J.L.S. (Kir3.4*, Kir2.1), Q.Z. (Kir4.1/Kir5.1) and R.R. (Kir3.1/Kir3.2, Kir4.1) performed the inside-out macropatch recordings. A.A.R.-M. carried out the PtdIns(4,5)P2 dose-response curve and Z.Z. performed the Na+ dose-response curve. D.E.L. initiated and supervised the work, edited the manuscript produced by A.R.-D., and revised the final form of the manuscript.

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Correspondence to Diomedes E Logothetis.

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Rosenhouse-Dantsker, A., Sui, J., Zhao, Q. et al. A sodium-mediated structural switch that controls the sensitivity of Kir channels to PtdIns(4,5)P2. Nat Chem Biol 4, 624–631 (2008). https://doi.org/10.1038/nchembio.112

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