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Activation of inwardly rectifying K+ channels by distinct PtdIns(4,5)P2 interactions

Nature Cell Biology volume 1, pages 183–188 (1999)Cite this article

Abstract

Direct interactions of phosphatidylinositol-4,5-bisphosphate (PtdIns(4,5)P2) with inwardly rectifying potassium channels are stronger with channels rendered constitutively active by binding to PtdIns(4,5)P2, such as IRK1, than with G-protein-gated channels (GIRKs). As a result, PtdIns(4,5)P2 alone can activate IRK1 but not GIRKs, which require extra gating molecules such as the βγ subunits of G proteins or sodium ions. Here we identify two conserved residues near the inner-membrane interface of these channels that are critical in interactions with PtdIns(4,5)P2. Between these two arginines, a conservative change of isoleucine residue 229 in GIRK4 to the corresponding leucine found in IRK1 strengthens GIRK4–PtdIns(4,5)P2 interactions, eliminating the need for extra gating molecules. A negatively charged GIRK4 residue, two positions away from the most strongly interacting arginine, mediates stimulation of channel activity by sodium by strengthening channel–PtdIns(4,5)P2 interactions. Our results provide a mechanistic framework for understanding how distinct gating mechanisms of inwardly rectifying potassium channels allow these channels to subserve their physiological roles.

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Figure 1: Identification of a region responsible for the differences in sensitivity of GIRK4 and IRK1 to PtdIns(4,5)P2.
Figure 2: Identification of amino-acid differences between GIRK4 and IRK1 that contribute to their differential sensitivities to PtdIns(4,5)P2.
Figure 3: Single point mutations in GIRK4 strengthen interactions with PtdIns(4,5)P2 and alleviate the absolute requirement of gating molecules for activation.
Figure 4: Gβγ stabilizes interactions of both GIRK4* and GIRK4*(I229L) with PtdIns(4,5)P2.
Figure 5: Binding of Na+ ions at a negatively charged residue modulates the strength of channel interactions with PtdIns(4,5)P2.

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Acknowledgements

We thank J.C. Garrison for purified Gβ1γ7 subunits; L. Lontsman for oocyte preparation; and R.F. Margolskee, S. Kupfer and M.M. Zhou for critical comments on the manuscript. This work was supported by a grant from the NIH (HL59949). H.Z. was supported by the Howard Hughes Medical Institute. T.M. was supported by an NIH training grant (HL07824).

Correspondence and requests for materials should be addressed to D.E.L.

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  1. Department of Physiology and Biophysics, Howard Hughes Medical Institute, Mount Sinai School of Medicine, CUNY, New York,, 10029, New York, USA

    Hailin Zhang

  2. Department of Physiology and Biophysics, Mount Sinai School of Medicine, CUNY, New York,, 10029,, New York, USA

    Cheng He, Xixin Yan, Tooraj Mirshahi & Diomedes E. Logothetis

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Zhang, H., He, C., Yan, X. et al. Activation of inwardly rectifying K+ channels by distinct PtdIns(4,5)P2 interactions. Nat Cell Biol 1, 183–188 (1999). https://doi.org/10.1038/11103

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