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Cytoplasmic domain structures of Kir2.1 and Kir3.1 show sites for modulating gating and rectification

Nature Neuroscience volume 8, pages 279–287 (2005)Cite this article

An Erratum to this article was published on 01 June 2005

Abstract

N- and C-terminal cytoplasmic domains of inwardly rectifying K (Kir) channels control the ion-permeation pathway through diverse interactions with small molecules and protein ligands in the cytoplasm. Two new crystal structures of the cytoplasmic domains of Kir2.1 (Kir2.1L) and the G protein–sensitive Kir3.1 (Kir3.1S) channels in the absence of PIP2 show the cytoplasmic ion-permeation pathways occluded by four cytoplasmic loops that form a girdle around the central pore (G-loop). Significant flexibility of the pore-facing G-loop of Kir2.1L and Kir3.1S suggests a possible role as a diffusion barrier between cytoplasmic and transmembrane pores. Consistent with this, mutations of the G-loop disrupted gating or inward rectification. Structural comparison shows a di-aspartate cluster on the distal end of the cytoplasmic pore of Kir2.1L that is important for modulating inward rectification. Taken together, these results suggest the cytoplasmic domains of Kir channels undergo structural changes to modulate gating and inward rectification.

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Figure 1: Sequence alignment and size-exclusion chromatograms of Kir3.1S, Kir2.1L, and KirBac1.1 with secondary structure elements noted.
Figure 2: Structures of Kir3.1S and Kir2.1L.
Figure 3: Kir3.1S and Kir2.1L in comparison to KirBac1.1.
Figure 4: Mutations in G-loop disrupt gating and inward rectification.
Figure 5: Substitution of glycine at Ala306 of Kir2.1 does not alter PIP2 affinity.
Figure 6: Mutation of di-aspartate cluster in Kir2.1 changes inward rectification.

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Acknowledgements

We thank D. Clapham, M. Lazdunski and S. Hebert for GIRK4, GIRK2 and ROMK1 cDNAs, respectively. We also thank D. Kaiser for analytical ultracentrifugation, C. Park for mass spectroscopy, and the staff at ALS and SSRL for X-ray data collection. This work was supported by grants from the National Institutes of Health (P.A.S & S.C.) and the McKnight Endowment for Neuroscience (P.A.S). S.C. acknowledges the support from the American Heart Association.

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  1. Structural Biology, The Salk Institute, La Jolla, 92037, California, USA

    Scott Pegan, Wei Zhou, Witek Kwiatkowski & Senyon Choe

  2. Peptide Biology Laboratories, The Salk Institute, La Jolla, 92037, California, USA

    Christine Arrabit & Paul A Slesinger

  3. Department of Pharmaceutical Sciences, College of Pharmacy, Oregon State University, Corvallis, 97331, Oregon, USA

    Anthony Collins

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Correspondence to Paul A Slesinger or Senyon Choe.

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Supplementary Fig. 1

Size chromatograms of Kir3.1S, Kir2.1L, and Kir3.2's N- and C-terminal domains expressed dicistronically. (PDF 209 kb)

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Pegan, S., Arrabit, C., Zhou, W. et al. Cytoplasmic domain structures of Kir2.1 and Kir3.1 show sites for modulating gating and rectification. Nat Neurosci 8, 279–287 (2005). https://doi.org/10.1038/nn1411

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