Selective Expression in Carotid Body Type I Cells of a Single Splice Variant of the Large Conductance Calcium- and Voltage-activated Potassium Channel Confers Regulation by AMP-activated Protein Kinase

Fiona A. Ross, J. Nicole Rafferty, Mark L. Dallas, Oluseye Ogunbayo, Naoko Ikematsu, Heather McClafferty, Lijun Tian, Helene Widmer, Iain C. M. Rowe, Christopher N. Wyatt, Michael J. Shipston, Chris Peers, D. Grahame Hardie, A. Mark Evans

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    34 Citations (Scopus)

    Abstract

    Inhibition of large conductance calcium-activated potassium (BKCa) channels mediates, in part, oxygen sensing by carotid body type I cells. However, BKCa channels remain active in cells that do not serve to monitor oxygen supply. Using a novel, bacterially derived AMP-activated protein kinase (AMPK), we show that AMPK phosphorylates and inhibits BKCa channels in a splice variant-specific manner. Inclusion of the stress-regulated exon within BKCa channel alpha subunits increased the stoichiometry of phosphorylation by AMPK when compared with channels lacking this exon. Surprisingly, however, the increased phosphorylation conferred by the stress-regulated exon abolished BKCa channel inhibition by AMPK. Point mutation of a single serine (Ser-657) within this exon reduced channel phosphorylation and restored channel inhibition by AMPK. Significantly, RT-PCR showed that rat carotid body type I cells express only the variant of BKCa that lacks the stress-regulated exon, and intracellular dialysis of bacterially expressed AMPK markedly attenuated BKCa currents in these cells. Conditional regulation of BKCa channel splice variants by AMPK may therefore determine the response of carotid body type I cells to hypoxia.

    Original languageEnglish
    Pages (from-to)11929-11936
    Number of pages8
    JournalJournal of Biological Chemistry
    Volume286
    Issue number14
    DOIs
    Publication statusPublished - 8 Apr 2011

    Keywords

    • CA2+-ACTIVATED K+ CURRENTS
    • PHOSPHORYLATION
    • HYPOXIA
    • INHIBITION
    • SENSITIVITY
    • MECHANISM
    • SUBSTRATE
    • ENERGY
    • ALPHA
    • CA

    Cite this

    Ross, F. A., Rafferty, J. N., Dallas, M. L., Ogunbayo, O., Ikematsu, N., McClafferty, H., Tian, L., Widmer, H., Rowe, I. C. M., Wyatt, C. N., Shipston, M. J., Peers, C., Hardie, D. G., & Evans, A. M. (2011). Selective Expression in Carotid Body Type I Cells of a Single Splice Variant of the Large Conductance Calcium- and Voltage-activated Potassium Channel Confers Regulation by AMP-activated Protein Kinase. Journal of Biological Chemistry, 286(14), 11929-11936. https://doi.org/10.1074/jbc.M110.189779