STIM1 gates the store-operated calcium channel ORAI1 in vitro

Yubin Zhou, Paul Meraner, Hyoung T. Kwon, Danya Machnes, Masatsugu Oh-Hora, Jochen Zimmer, Yun Huang, Antonio Stura, Anjana Rao, Patrick G. Hogan

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    Abstract

    Store-operated Ca2+ entry through the plasma membrane Ca2+ release–activated Ca2+ (CRAC) channel in mammalian T cells and mast cells depends on the sensor protein stromal interaction molecule 1 (STIM1) and the channel subunit ORAI1. To study STIM1-ORAI1 signaling in vitro, we have expressed human ORAI1 in a sec6-4 strain of the yeast Saccharomyces cerevisiae and isolated sealed membrane vesicles carrying ORAI1 from the Golgi compartment to the plasma membrane. We show by in vitro Ca2+ flux assays that bacterially expressed recombinant STIM1 opens wild-type ORAI1 channels but not channels assembled from the ORAI1 pore mutant E106Q or the ORAI1 severe combined immunodeficiency (SCID) mutant R91W. These experiments show that the STIM1-ORAI1 interaction is sufficient to gate recombinant human ORAI1 channels in the absence of other proteins of the human ORAI1 channel complex, and they set the stage for further biochemical and biophysical dissection of ORAI1 channel gating.

    Original languageEnglish
    Pages (from-to)112-117
    Number of pages6
    JournalNature Structural and Molecular Biology
    Volume17
    Issue number1
    DOIs
    Publication statusPublished - Jan 2010

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    All Science Journal Classification (ASJC) codes

    • Structural Biology
    • Molecular Biology

    Cite this

    Zhou, Y., Meraner, P., Kwon, H. T., Machnes, D., Oh-Hora, M., Zimmer, J., ... Hogan, P. G. (2010). STIM1 gates the store-operated calcium channel ORAI1 in vitro. Nature Structural and Molecular Biology, 17(1), 112-117. https://doi.org/10.1038/nsmb.1724